Compounds for modulating TRPV3 function

ABSTRACT

The present application relates to compounds and methods for treating pain and other conditions related to TRPV3.

RELATED APPLICATIONS

This application claims priority to U.S. provisional application Ser.No. 60/679,438, filed May 9, 2005, U.S. provisional application Ser. No.60/679,436, filed May 9, 2005, and U.S. provisional application Ser. No.60/702,584, filed Jul. 25, 2005. The disclosures of each of theforegoing applications are hereby incorporated by reference in theirentirety.

BACKGROUND

A variety of ion channel proteins exist to mediate ion flux acrosscellular membranes. The proper expression and function of ion channelproteins is essential for the maintenance of cell function,intracellular communication, and the like. Numerous diseases are theresult of misregulation of membrane potential or aberrant calciumhandling. Given the central importance of ion channels in modulatingmembrane potential and ion flux in cells, identification of agents thatcan promote or inhibit particular ion channels are of great interest asresearch tools and as possible therapeutic agents.

One such channel is the Transient Receptor Potential V3 (TRPV3) channel.TRPV3 is a calcium permeable channel, specifically a calcium permeablenon-selective cation channel. In addition to calcium ions, TRPV3channels are permeable to other cations, for example sodium. Thus, TRPV3channels modulate membrane potential by modulating the flux of cationssuch as calcium and sodium ions. Although non-selective cation channelssuch as TRPV3 modulate, among other things, calcium ion flux, they aremechanistically distinct from voltage-gated calcium channels. Generally,voltage-gated calcium channels respond to membrane depolarization andopen to permit an influx of calcium from the extracellular medium thatresults in an increase in intracellular calcium levels orconcentrations. In contrast, TRP channels which are non-selective cationchannels are generally signal transduction gated, long lasting, andproduce more prolonged changes in ion concentration. These mechanisticdifferences are accompanied by structural differences amongvoltage-gated and TRP channels. Thus, although many diverse channels actto regulate ion flux and membrane potential in various cell types and inresponse to numerous stimuli, it is important to recognize thesignificant structural, functional, and mechanistic differences amongdifferent classes of ion channels.

TRPV3 function has been implicated in, among other things, the receptionand transduction of pain. Accordingly, it would be desirable to identifyand make compounds that can modulate one or more functions of TRPV3.Such compounds have a variety of in vitro and in vivo uses.

SUMMARY

An important aspect of achieving cellular homeostasis is the maintenanceof appropriate ion concentrations in various cell types during thedevelopment of and in response to numerous stimuli. Large numbers ofdiverse types of ion channels act to maintain cellular homeostasis bymoving ions into and out of cells across the plasma membrane, and withincells by moving ions across membranes of intracellular organellesincluding, for example, the endoplasmic reticulum, sarcoplasmicreticulum, mitochondria and endocytic organelles including endosomes andlysosomes. One such ion channel is the non-selective cation channelTRPV3. TRPV3 is cation permeable and belongs to the larger family of TRPion channels.

TRP channels have been classified into at least six groups: TRPC(short), TRPV (vanilloid), TRPM (long, melastatin), TRPP (polycystins),TRPML (mucolipins), and TRPA (ANKTM1). The TRPC group can be dividedinto 4 subfamilies (TRPC1, TRPC4,5, TRPC3,6,7 and TRPC2) based onsequence homology and functional similarities. Currently the TRPV familyhas 6 members. TRPV5 and TRPV6 are more closely related to each otherthan to TRPV1, TRPV2, TRPV3, or TRPV4. TRPV3 is most closely related toTRPV4, and is more closely related to TRPV1 and TRPV2 than to TRPV5 andTRPV6. The TRPM family has 8 members. Constituents include thefollowing: the founding member TRPM1 (Melastatin or LTRPC1), TRPM3(KIAA1616 or LTRPC3), TRPM7 (TRP-PLIK, ChaK(1), LTRPC7), TRPM6 (ChaK2),TRPM2 (TRPC7 or LTRPC2), TRPM8 (Trp-p8 or CMR1), TRPM5 (Mtr1 or LTRPC5),and TRPM4 (FLJ20041 or LTRPC4). The sole mammalian member of the TRPAfamily is ANKTM1. The TRPML family consists of the mucolipins, whichinclude TRPML1 (mucolipins 1), TRPML2 (mucolipins 2), and TRPML3(mucolipin3). The TRPP family consists of two groups of channels: thosepredicted to have six transmembrane domains and those that have 11.TRPP2 (PKD2), TRPP3 (PKD2L1), TRPP5 (PKD2L2) are all predicted to havesix transmembrane domains. TRPP1 (PKD1, PC1), PKD-REJ and PKD-1L1 areall thought to have 11 transmembrane domains.

The TRP channels constitute a large and important class of channelsinvolved in modulating cellular homeostasis. The present inventionprovides methods and compositions that modulate at least one TRP familymember. Specifically, the present invention provides methods andcompositions for antagonizing a function of TRPV3. Modulating a functionof TRPV3 provides a means for modulating calcium homeostasis, sodiumhomeostasis, intracellular calcium levels, membrane polarization(resting membrane potential), and/or cation levels in a cell. Compoundsthat can modulate one or more TRPV3 functions are useful in many aspectsincluding, but not limited to, maintaining calcium homeostasis;maintaining sodium homeostasis; modulating intracellular calcium levels;modulating membrane polarization (membrane potential); modulating cationlevels; and/or treating or preventing diseases, disorders, or conditionsassociated with calcium homeostasis, sodium homeostasis, calcium orsodium dyshomeostasis, or membrane polarization/hyperpolarization(including hypo and hyperexcitability), and/or treating or preventingdiseases, disorders, or conditions associated with regulation ormisregulation of TRPV3 expression or function. Additionally, the presentinvention provides, in certain embodiments, methods and compositionsthat antagonize both a function of TRPV3 and a function of one or moreadditional TRP channels.

The present application provides compounds that can modulate TRPV3function. Methods employing these compounds are also provided. Certainembodiments provide a method of modulating a TRPV3 function in a cellcomprising administering to the cell an effective amount of a compoundthat inhibits a TRPV3 mediated current. Certain embodiments provide amethod of modulating a TRPV3 function in a cell comprising administeringto the cell an effective amount of a compound that inhibits TRPV3function, wherein the compound inhibits the Phase II outward currentmediated by TRPV3. Certain embodiments provide a method of preventing ortreating a disease or condition related to TRPV3 function in a subjectcomprising administering to the subject a therapeutically effectiveamount of a compound that inhibits TRPV3 function, wherein the compoundinhibits the Phase II outward current mediated by TRPV3. Certainembodiments provide a method of modulating a TRPV3 function in a cellcomprising administering to the cell an effective amount of a compoundthat inhibits TRPV3 function, wherein the compound inhibits the Phase IIinward current mediated by TRPV3. Certain embodiments also provide amethod of preventing or treating a disease or condition related to TRPV3function in a subject comprising administering to the subject atherapeutically effective amount of a compound that inhibits TRPV3function, wherein the compound inhibits the Phase II inward currentmediated by TRPV3. Certain embodiments provide a method of modulatingTRPV3 function in a cell comprising administering to the cell aneffective amount of a compound that inhibits TRPV3 function, wherein thecompound inhibits the Phase I inward current mediated by TRPV3. Certainembodiments also provide a method of preventing or treating a disease orcondition related to TRPV3 function in a subject comprisingadministering to the subject a therapeutically effective amount of acompound that inhibits TRPV3 function, wherein the compound inhibits thePhase I inward current mediated by TRPV3. Certain embodiments alsoprovide a method of preventing or treating a disease or conditionrelated to TRPV3 function in a subject comprising administering to thesubject a therapeutically effective amount of a compound that inhibitsTRPV3 function, wherein the compound inhibits the Phase I outwardcurrent mediated by TRPV3. Certain embodiments provide a method ofmodulating TRPV3 function in a cell comprising administering to the cellan effective amount of a compound that inhibits TRPV3 function, whereinthe compound inhibits the Phase I outward current mediated by TRPV3.Certain embodiments also provide a method of preventing or treating adisease or condition involving activation of TRPV3 or for which reducedTRPV3 activity can reduce the severity in a subject comprisingadministering to the subject a therapeutically effective amount of acompound that inhibits TRPV3 function, wherein the compound inhibits oneor more of a Phase I inward current mediated by TRPV3, a Phase II inwardcurrent mediated by TRPV3, a Phase I outward current mediated by TRPV3,or a Phase II outward current mediated by TRPV3. In any of theforegoing, the invention additionally provides compounds and methodsthat inhibit both the Phase I outward current and the Phase II outwardcurrent. Furthermore, in any of the foregoing, the invention providescompounds and methods that inhibit both the Phase I inward current andthe Phase II inward current, as well as compounds that inhibit anycombination of Phase I and Phase II currents. Note that inhibition of aparticular current refers to the ability of a compound to inhibit thatcurrent (e.g., Phase I inward, Phase I outward, Phase II inward, and/orPhase II outward) in either an in vitro or in vivo assay. Inhibition ofa particular current in either an in vivo or an in vitro assay serves asa proxy for the particular functional activity of the particularcompound.

The biphasic currents mediated by TRPV3 are discussed in, for example,Chung et al. (Chung et al., 2005, Journal of Biological Chemistry 280:15928-15941). Briefly, a unique property of TRPV3 is that there is aphase change in the current. The current-voltage relationship changesupon repeated stimulation, so that the amount of inward currentincreases dramatically. For ease, two phases of TRPV3 current have beendescribed: Phase I and Phase II. Throughout, we have defined phase I ascurrents that show a 10:1 ratio or greater of outward current amplitude(at +100 mV) to inward current amplitude (at −120 mV). In other words,the current shows strong outward rectification and minimal inwardcurrent. Phase II is defined as a ration of 2:1 or less of outwardcurrent amplitude (at +100 mV) to inward current amplitude (at −120 mV).The current-voltage relationship is fairly linear in this case.

The following articles are exemplary of the state of the art regardingthe structure and function of TRPV3 (Ramsey et al. (2006) Annual RevPhysiology 68: 619-647; Clapham. (2003) Nature 426: 517-524; Xu et al.(2002) Nature 418: 181-186; Clapham et al. (2001) Nature Reviews ofNeuroscience 2: 387-396). The foregoing articles are incorporated byreference in their entirety.

One aspect of the present invention relates to a method for treating orpreventing a condition involving activation of TRPV3 or for whichreduced TRPV3 activity can reduce the severity by administering a TRPV3antagonist that inhibits TRPV3-mediated current. Described in greaterdetail below are TRPV3 antagonists that have measured IC₅₀'s forinhibition of TRPV3 of 10 micromolar or less, 1 micromolar or less, 500nanomolar or less, 200 nanomolar or less, 100 nanomolar or less, andeven 10 nanomolar or less. In certain embodiments, the TRPV3 antagonistinhibit one or both of inward and outward TRPV3-mediated current with anIC₅₀ of 1 micromolar or less, and more with an IC₅₀ of 500 nanomolar orless, 200 nanomolar or less, 100 nanomolar or less, 25 nanomolar or lessand even 10 nanomolar or less. In certain embodiments, the TRPV3antagonist inhibits at least 95% of TRPV3-mediated current at 5micromolar or less, and even more preferably at 1 micromolar or less.

In certain embodiments, the subject TRPV3 antagonists inhibit TRPV3 withan IC₅₀ at least one order of magnitude lower than its IC₅₀ forinhibition of one or more of TRPV5, TRPV6, NaV 1.2, TRPV1, mitochondrialuniporter and hERG channel activities, and even more preferably two oreven three orders of magnitude lower.

In certain embodiments, the TRPV3 antagonists inhibit TRPV3 with an IC₅₀at least one order of magnitude lower than its IC₅₀ for inhibition ofTRPV1, and even more preferably two or even three orders of magnitudelower. In certain embodiments, the subject TRPV3 antagonists can beselected for selectivity for TRPV3 versus TRPV1 on the basis of havingIC₅₀ for TRPV1 inhibition greater than 10 micromolar.

In certain embodiments, the TRPV3 antagonists inhibit one or more ofTRPV2, TRPV4, ANKTM1 and/or TRPM8 with an IC₅₀ of 10 micromolar or less.

In certain embodiments, the TRPV3 antagonist has a therapeutic index(T.I.) for treating the condition with the compound of 10 or greater,and even more preferably has a T.I. of at least 25, 50 or even 100.

In preferred embodiments, the TRPV3 inhibitor has an IC₅₀ for TRPV3inhibition that, at that concentration, does not cause QT intervalelongation in the patient nor alter temperature regulation in thepatient.

In certain embodiments, the TRPV3 inhibitor is used to treat orameliorate pain. Exemplary classes of pain that can treated using aTRPV3 inhibitor include, but are not limited to nociceptive pain,inflammatory pain, and neuropathic pain. Pain that can be treated with aTRPV3 inhibitor can be chronic or acute.

In certain embodiments, the TRPV3 inhibitor is non-narcotic and haslittle or no narcotic side-effects. In certain other embodiments, theTRPV3 inhibitor can be used to treat or ameliorate pain with fewerside-effects than narcotic pain relievers. Exemplary side-effects thatmay be substantially absent at effective dosages of TRPV3 inhibitorsinclude one or more of exopthalmos, catalepsy, disruption of gutmotility, and inhibiton of sensation in non-injured areas of the body.

In certain embodiments, the TRPV3 antagonist is “small molecule”, e.g.,an organic molecule having a molecular weight of 2000 amu or less.Exemplary TRPV3 antagonists include a compound of Formula I or a salt,solvate, hydrate, oxidative metabolite or prodrug thereof:

wherein: Ar represents an aryl or heteroaryl group; Y represents Ph,OArl, SArl, or N(U)(Arl); R represents H or a lower alkyl; X representsCH₂, O, S, NR₁₀, CF₂, or C(CN)₂; W represents O, S, or NR′; n is 1, orwhen X is CH₂, n is 1 or 2; Arl represents an aralkyl, heteroaralkyl,aryl, or heteroaryl group, which may be monocyclic or bicyclic; Phrepresents a substituted or unsubstituted phenyl ring, optionally fusedto a substituted or unsubstituted 5- to 7-membered cycloalkyl,heterocyclyl, aryl, or heteroaryl ring; R′, independently for eachoccurrence, represents H or a lower alkyl; U represents hydrogen orlower alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl,heteroaryl, cycloalkylalkyl, heterocyclylalkyl, aralkyl, orheteroaralkyl, or, U, N and R′ taken together form a 5- to 7-memberedheterocyclic ring, or, U, Arl and N taken together form a ring fused tothe aryl or heteroaryl ring of Arl, thereby forming a bicyclicstructure; R₁₀ represents H, lower alkyl, or cyano; and wherein saidcompound inhibits TRPV3 with an with an IC₅₀ of 10 micromolar or less.

In certain embodiments, R is H, W is O or S, and X is O or S.

In certain embodiments, X is CH₂.

In certain preferred embodiments, the TRPV3 antagonist is represented inFormula Ia, or is a salt, solvate, hydrate, oxidative metabolite orprodrug thereof:

wherein: Ar represents an aryl or heteroaryl group; Y represents Ph,OArl, SArl, or N(U)(Arl); R represents H or a lower alkyl; X representsO, S, NR₁₀, CF₂, or C(CN)₂; W represents O, S, or NR′; Arl represents anaralkyl, heteroaralkyl, aryl, or heteroaryl group, which may bemonocyclic or bicyclic; Ph represents a substituted or unsubstitutedphenyl ring, optionally fused to a substituted or unsubstituted 5- to7-membered cycloalkyl, heterocyclyl, aryl, or heteroaryl ring; R′,independently for each occurrence, represents H or a lower alkyl; Urepresents hydrogen or lower alkyl, alkenyl, alkynyl, cycloalkyl,heterocyclyl, aryl, heteroaryl, cycloalkylalkyl, heterocyclylalkyl,aralkyl, or heteroaralkyl, or, U, N and R′ taken together form a 5- to7-membered heterocyclic ring, or, U, Arl and N taken together form aring fused to the aryl or heteroaryl ring of Arl, thereby forming abicyclic structure; and R₁₀ represents H, lower alkyl, or cyano.

In certain embodiments of Formulas I and Ia, Y is N(U)(Arl), and in suchembodiments, Y can represent, for example,

wherein: R₁ and R₂, each independently, is absent or represents one ormore substituents on the ring to which it is attached; Ar′ represents anaryl or heteroaryl group; R₅ represents CH₂, O, NR₆; R₆ represents H orlower alkyl; and n represents 1, 2, or 3. In certain embodiments of theabove-referenced compounds, one or more of the following conditions maybe true:

-   -   Ar′ represents phenyl, thiophene, imidazole, thiazole, furan,        oxazole, tetrazole, thiadiazole, pyridine, pyrimadine, or        triazole;    -   each R₁ is independently selected from H, lower alkyl, alkoxy,        carboxyl, ester, amido, sulfonamido, heterocyclyl, cycloalkyl,        hydroxyl, amino, acylamino, thioether, sulfonylamino, or R₁        taken together with the carbon to which it is attached forms a        carbonyl or thiocarbonyl;    -   each R2 is independently selected from H, lower alkyl, alkoxy,        carboxyl, ester amido, sulfonamido, heterocyclyl, cycloalkyl,        hydroxyl, amino, acylamino, thioether, sulfonylamino, halogen,        CF₃, or cyano;

In certain preferred embodiments of Formulas I and Ia, Y represents

wherein R₁ and R₂ each, independently, are absent or represent one ormore substituents on the ring to which they are attached. Even morepreferably, independently for each occurrence, R₁ is selected from H,lower alkyl, alkoxy, carboxyl, ester, amido, sulfonamido, heterocyclyl,cycloalkyl, hydroxyl, amino, acylamino, thioether, sulfonylamino, or R1taken together with the carbon to which it is attached forms a carbonylor thiocarbonyl; and independently for each occurrence, R₂ is selectedfrom H, lower alkyl, alkoxy, carboxyl, ester, amido, sulfonamido,heterocyclyl, cycloalkyl, hydroxyl, amino, acylamino, thioether,sulfonylamino, halogen, CF₃, or cyano.

For further illustration, Y can represent

wherein R₁ represents H, lower alkyl, alkoxy, carboxyl, ester, amido,sulfonamido, heterocyclyl, cycloalkyl, hydroxyl, amino, acylamino,thioether, sulfonylamino, or R₁ taken together with the carbon to whichit is attached forms a carbonyl or thiocarbonyl; and R₂ represents H,lower alkyl, alkoxy, carboxyl, ester, amido, sulfonamido, heterocyclyl,cycloalkyl, hydroxyl, amino, acylamino, thioether, sulfonylamino,halogen, CF₃, or cyano. In certain preferred embodiments, R₁ representsH, lower alkyl, alkoxy, or R₁ taken together with the carbon to which itis attached forms a carbonyl, and R₂ represents H, lower alkyl, alkoxyor halogen.

In certain preferred embodiments of Formulas I and Ia, Ar represents

wherein: Cy represents an aryl or heteroaryl ring; Z represents O, S,NR₁₁, CF₂, or C(CN)₂; R₃ is absent or represents one or moresubstituents on the ring to which it is attached; R₁₁ represents H,lower alkyl, or cyano; A represents N or C(R_(2a)); and R_(2a)represents H, lower alkyl, alkoxy, or halogen.

For example, Cy can represent a phenyl, thiophene, imidazole, thiazole,furan, oxazole, tetrazole, thiadiazole, pyridine, pyrimadine, ortriazole, and R₃ can be absent, or independently for each occurrence,selected from lower alkyl, alkoxy, carboxyl, ester, amido, sulfonamido,heterocyclyl, cycloalkyl, hydroxyl, amino, acylamino, thioether,sulfonylamino, nitro, halogen, CF₃, or cyano. In certain preferredembodiments, A represents N, and Z represents O or S.

To further illustrate, Ar can represent

wherein: Z represents O, S, NR₁₁, CF₂, NR, or C(CN)₂; A represents N orC(R_(2a)); R₃ is absent or represents one or more substituents on thering to which it is attached; R₁₁ represents H, lower alkyl, or cyano;and R_(2a) represents H, lower alkyl, alkoxy, or halogen. In certainpreferred embodiments: Z represents O or S; and R₃ is absent or each R₃is independently selected from lower alkyl, alkoxy, carboxyl, ester,amido, sulfonamido, heterocyclyl, cycloalkyl, hydroxyl, amino,acylamino, thioether, sulfonylamino, nitro, halogen, CF₃, or cyano;

In certain preferred embodiments, Ar can represent

wherein: Z represents O, S, NR₁₁, CF₂, or C(CN)₂; A represents N orC(R_(2a)); R₃ independently for each occurrence represents H, loweralkyl, alkoxy, carboxyl, ester, amido, sulfonamido, heterocyclyl,cycloalkyl, hydroxyl, amino, acylamino, thioether, sulfonylamino,halogen, CF₃, or cyano; R₁₁ represents H, lower alkyl, or cyano; andR_(2a) represents H, lower alkyl, alkoxy, or halogen. Preferrably Zrepresents O or S, and R₃ independently for each occurrence representsH, lower alkyl, alkoxy, CF₃, nitro, or halogen.

In certain other preferred embodiments of Formulas I and Ia, Arrepresents

wherein: G′ represents N or C(R₅); R₃ is absent or represents one ormore substituents on Cy; R₅ represents H or a substituent; R₄ representsH or a substituent; and Cy represents an aryl or heteroaryl ring. Incertain preferred embodiments, Cy can be a phenyl, thiophene, imidazole,thiazole, furan, oxazole, tetrazole, thiadiazole, pyridine, pyrimadine,or triazole. In certain embodiments, R₃ is absent, or independently foreach occurrence, is selected from lower alkyl, alkoxy, carboxyl, ester,amido, sulfonamido, heterocyclyl, cycloalkyl, hydroxyl, amino,acylamino, thioether, sulfonylamino, nitro, halogen, CF₃, or cyano; andR₄ and R₅, independently for each occurrence, are selected from H, loweralkyl, alkoxy, carboxyl, ester, amido, sulfonamido, heterocyclyl,cycloalkyl, hydroxyl, amino, acylamino, thioether, sulfonylamino,halogen, CF₃, or cyano.

In still other embodiments, the TRPV3 antagonist is a compound ofFormula V or a salt, solvate, hydrate, oxidative metabolite or prodrugthereof:

wherein: X represents O, S, NR₁₀, CF₂, or C(CN)₂; Z represents O, S,NR₁₁, CF₂, or C(CN)₂; R₁₀ represents H, lower alkyl, or cyano; R₁₁represents H, lower alkyl, or cyano; A represents N or C(R₂); Grepresents N or C(R₃); R₁ is absent or represents one or moresubstituents on the ring to which it is attached; R₂ represents,independently for each occurrence, H or a substituent; R₃ represents,independently for each occurrence, H or a substituent; and D representsO or N.

In certain preferred embodiments of formula V: X represents S and Drepresents O; R₁ is absent, or independently for each occurrence, isselected from lower alkyl, alkoxy, carboxyl, ester, amido, sulfonamido,heterocyclyl, cycloalkyl, hydroxyl, amino, acylamino, thioether,sulfonylamino, or R₁ taken together with the carbon to which it isattached forms a carbonyl or thiocarbonyl; and R₂ and R₃, independentlyfor each occurrence, are selected from H, lower alkyl, alkoxy, carboxyl,ester, amido, sulfonamido, heterocyclyl, cycloalkyl, hydroxyl, amino,acylamino, thioether, sulfonylamino, nitro, halogen, CF₃, or cyano.

To further illustrate, the TRPV3 antagonist can be represented byFormula Va:

wherein: R₁, R₂ and R₃ are absent or represent one or more substituentson the ring to which they are attached. For instance, R₁ is absent, orindependently for each occurrence, is selected from lower alkyl, alkoxy,carboxyl, ester, amido, sulfonamido, heterocyclyl, cycloalkyl, hydroxyl,amino, acylamino, thioether, sulfonylamino, or R₁ taken together withthe carbon to which it is attached forms a carbonyl or thiocarbonyl; andR₂ and R₃ are absent, or independently for each occurrence, are selectedfrom lower alkyl, alkoxy, carboxyl, ester, amido, sulfonamido,heterocyclyl, cycloalkyl, hydroxyl, amino, acylamino, thioether,sulfonylamino, nitro, halogen, CF₃, or cyano. In certain preferredembodiments, R₁ is absent, or independently for each occurrence, isselected from lower alkyl, or alkoxy, or R₁ taken together with thecarbon to which it is attached forms a carbonyl; R₂ is absent, orindependently for each occurrence, is selected from lower alkyl, alkoxy,or halogen; and R₃ is absent, or independently for each occurrence, isselected from lower alkyl, alkoxy, CF₃, nitro, or halogen. In even morepreferred embodiments, R₁ represents H; R₂ represents H or CH₃ and ispositioned ortho to the N of the ring system; and R₃ represents one ormore substituents positioned meta or para to the nitrogen of the ringsystem.

In still another illustration, the TRPV3 antagonist can be representedby Formula Vb:

wherein R₁, R₂ and R₃ are absent or represent one or more substituentson the ring to which they are attached. For instance, R₁ is absent, orindependently for each occurrence, is selected from lower alkyl, alkoxy,carboxyl, ester, amido, sulfonamido, heterocyclyl, cycloalkyl, hydroxyl,amino, acylamino, thioether, sulfonylamino, or R₁ taken together withthe carbon to which it is attached forms a carbonyl or thiocarbonyl; andR₂ and R₃ are absent, or independently for each occurrence, are selectedfrom lower alkyl, alkoxy, carboxyl, ester, amido, sulfonamido,heterocyclyl, cycloalkyl, hydroxyl, amino, acylamino, thioether,sulfonylamino, nitro, halogen, CF₃, or cyano. In certain preferredembodiments, R₁ is absent, or independently for each occurrence, isselected from lower alkyl, or alkoxy, or R₁ taken together with thecarbon to which it is attached forms a carbonyl; R₂ is absent, orindependently for each occurrence, is selected from lower alkyl, alkoxy,or halogen; and R₃ is absent, or independently for each occurrence, isselected from lower alkyl, alkoxy, CF₃, nitro, or halogen. In certainembodiments, R₁, R₂, and R₃ are absent.

In still other embodiments, the TRPV3 antagonist is a compound ofFormula VI or a salt, solvate, hydrate, oxidative metabolite or prodrugthereof:

wherein: X represents O, S, NR₁₀, CF₂, or C(CN)₂; R₁₀ represents H,lower alkyl, or cyano; A represents N or C(R₂); G represents N or C(R₃);R₁ is absent or represents one or more substituents on the ring to whichit is attached; R₂ represents, independently for each occurrence, H or asubstituent; R₃ represents, independently for each occurrence, H or asubstituent; R₄ represents H or a substituent; D represents O or N; andwherein said compound inhibits TRPV3 with an with an IC₅₀ of 10micromolar or less.

In certain preferred embodiments, X represents S and D represents O.

In certain embodiments, the compound can be represented by Formula VIa:

wherein: R₁, R₂ and R₃ are absent or represent one or more substituentson the ring to which they are attached; and R₄ represents H or asubstituent. In certain preferred embodiments, R₁ is absent, orindependently for each occurrence, is selected from lower alkyl, alkoxy,carboxyl, ester, amido, sulfonamido, heterocyclyl, cycloalkyl, hydroxyl,amino, acylamino, thioether, sulfonylamino, or R₁ taken together withthe carbon to which it is attached forms a carbonyl or thiocarbonyl; andR₂, R₃ and R₄ are absent, or independently for each occurrence, isselected from H, lower alkyl, alkoxy, carboxyl, ester, amido,sulfonamido, heterocyclyl, cycloalkyl, hydroxyl, amino, acylamino,thioether, sulfonylamino, nitro, halogen, CF₃, or cyano. In preferredembodiments, the R₁ is absent, or independently for each occurrence, isselected from lower alkyl, or alkoxy, or R₁ taken together with thecarbon to which it is attached forms a carbonyl; R₂ is absent, orindependently for each occurrence, is selected from lower alkyl, alkoxy,or halogen; R₃ absent, or independently for each occurrence, is selectedfrom H, lower alkyl, alkoxy, CF₃, nitro, or halogen; and R₄ is selectedfrom H, lower alkyl, or cyano. In other preferred embodiments, R₁ and R₂are absent; R₃ represents methoxy positioned para to the N of the ringsystem; and R₄ represents cyano.

In certain embodiments, the compound can be represented by Formula VIb:

wherein: R₁, R₂ and R₃ are absent or represent one or more substituentson the ring to which they are attached; and R₄ represents H or asubstituent. For instance, R₁ is absent, or independently for eachoccurrence, is selected from lower alkyl, alkoxy, carboxyl, ester,amido, sulfonamido, heterocyclyl, cycloalkyl, hydroxyl, amino,acylamino, thioether, sulfonylamino, or R₁ taken together with thecarbon to which it is attached forms a carbonyl or thiocarbonyl; and R₂,R₃ and R₄ are absent, or independently for each occurrence, is selectedfrom lower alkyl, alkoxy, carboxyl, ester, amido, sulfonamido,heterocyclyl, cycloalkyl, hydroxyl, amino, acylamino, thioether,sulfonylamino, nitro, halogen, CF₃, or cyano. In certain preferredembodiments, R₁ is absent, or independently for each occurrence, isselected from lower alkyl, alkoxy, or halogen or R₁ taken together withthe carbon to which it is attached forms a carbonyl; R₂ is absent, orindependently for each occurrence, is selected from lower alkyl, alkoxy,or halogen; R₃ is absent, or independently for each occurrence, isselected from lower alkyl, alkoxy, CF₃, nitro, or halogen; and R₄ isselected from H, lower alkyl, or cyano.

In still other illustrative embodiments, the compound can be representedby Formula II or a salt, solvate, hydrate, oxidative metabolite orprodrug thereof:

wherein: W represents O or S; X′ represents NR₁₂; R₁₂ represents H orlower alkyl; Ar and Ar′ independently represent substituted orunsubstituted aryl or heteroaryl rings; and wherein said compoundinhibits TRPV3 with an with an IC₅₀ of 10 micromolar or less. Forinstance, Ar can represents phenyl, as can Ar′.

In yet another illustrative embodiments, the compound can be representedby Formula III or a salt, solvate, hydrate, oxidative metabolite orprodrug thereof:

wherein: Ar represents a substituted or unsubstituted aryl or heteroarylring; Q represents O, S, NR, or CH₂; R, independently for eachoccurrence, represents H or lower alkyl; R′ represents H or asubstituted or unsubstituted lower alkyl, alkenyl, alkynyl, cycloalkyl,heterocyclyl, aryl, heteroaryl, cycloalkylalkyl, heterocyclylalkyl,aralkyl, or heteroaralkyl; R″, independently for each occurrence,represents H or lower alkyl; V is absent or represents C═O, C═S, or SO₂;W represents a hydrogen-bond acceptor, such as ═O, ═S, amino, hydroxyl,or ether; E represents H, a substituted or unsubstituted lower alkyl,alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl,cycloalkylalkyl, heterocyclylalkyl, aralkyl, or heteroaralkyl, orhalogen, carbonyl (e.g., ester, carboxyl, or formyl), thiocarbonyl(e.g., thioester, thiocarboxylate, or thioformate), ketone, aldehyde,amino, acylamino, amido, amidino, cyano, nitro, azido, sulfonyl,sulfoxido, sulfate, sulfonate, sulfamoyl, sulfonamido, or phosphoryl; xrepresents 0 or 1 wherein when x is 0, the carbons bearing W and V aredirectly linked by a single or double bond; and wherein said compoundinhibits TRPV3 with an with an IC₅₀ of 10 micromolar or less.

In certain embodiments, the compounds of Formula III are characterizedby one or more of the following: Q represents S or CH₂; R represents H;R′ represents a substituted or unsubstituted aryl or heteroaryl ring; R″represents lower alkyl; and/or E represents CO₂Me or CO₂Et.

Additional illustrative embodiments of TRPV3 antagonists includecompounds can be represented by Formula IV or a salt, solvate, hydrate,oxidative metabolite or prodrug thereof:

wherein: Ar and Ar′ independently represent substituted or unsubstitutedaryl or heteroaryl rings; R, independently for each occurrence,represents H or lower alkyl; X, independently for each occurrence,represents O, S, NR, CF₂, or CR₂; M, independently for each occurrence,represents a substituted or unsubstituted methylene group; and whereinsaid compound inhibits TRPV3 with an with an IC₅₀ of 10 micromolar orless. For instance

In certain embodiments, the compounds of Formula IV are characterized byone or more of the following: Ar and Ar′ represent phenyl or pyridylrings; R represents H; X represents O or S; M represents CH₂.

The TRPV3 antagonist of the subject invention can be used as part of aprophylaxis or treatment for a variety of disorders and conditions,including, but not limited to, acute and/or chronic pain, touchsensitivity, burns, inflammation, diabetic neuropathy, psoriasis,eczema, dermatitis, post-herpetic neuralgia (shingles), migraine,incontinence, fever, hot flashes, osteoarthritis, oral mucositis, cancerpain, bladder cystits, pain associated with Crohn's disease andIrritable Bowel Syndrome (IBS), rheumatoid arthritis, Grierson-Gopalansyndrome (better known as burning feet syndrome), burning mouth syndrome(BMS) and cough, or is used as a depilatory to promote loss of orinhibit the growth of hair on a patient.

The TRPV3 antagonists can be administered alone or in combination withother therapeutic agents. For instance, the TRPV3 antagonists isadministered conjointly with one more of an anti-inflammatory agent,anti-acne agent, anti-wrinkle agent, anti-scarring agent, anti-psoriaticagent, anti-proliferative agent, anti-fungal agent, anti-viral agent,anti-septic agent, anti-migraine agent, keratolytic agent, or a hairgrowth inhibitor.

The TRPV3 antagonists can be administered topically, orally,transdermally, rectally, vaginally, parenterally, intranasally,intraocularly, intravenously, intramuscularly, intraarterially,intrathecally, intracapsularly, intraorbitally, intracardiacly,intradermally, intraperitoneally, transtracheally, subcutaneously,subcuticularly, intraarticularly, subcapsularly, subarachnoidly,intraspinally, intrasternally or by inhallation.

In certain preferred embodiments, the TRPV3 antagonists is administeredtopically.

In certain preferred embodiments, the TRPV3 antagonists is administeredorally.

In certain preferred embodiments, the TRPV3 antagonists is administeredparenterally.

In certain preferred embodiments, the TRPV3 antagonists is administeredto prevent, treat or alleviate signs and symptoms of acute pain, chronicpain, touch sensitivity, itching sensitivity, or as part of treating aburn, such as, for example, post-surgical pain, cancer pain, orneuropathic pain.

In certain preferred embodiments, the TRPV3 antagonists is administeredto prevent, treat or alleviate signs and symptoms of migraine.

In certain preferred embodiments, the TRPV3 antagonists is administeredto prevent, treat or alleviate signs and symptoms of a disorder orcondition selected from the group consisting of diabetic neuropathy,inflammation, psoriasis, eczema, dermatitis, post-herpetic neuralgia(shingles), incontinence, bladder incontinence, fever, hot flashes, andcough.

In certain preferred embodiments, the TRPV3 antagonists is administeredto prevent, treat or alleviate signs and symptoms of osteoarthritis.

In certain preferred embodiments, the TRPV3 antagonists is administeredto prevent, treat or alleviate signs and symptoms of rheumatoidarthritis.

In certain preferred embodiments, the TRPV3 antagonists is administeredto prevent, treat or alleviate signs and symptoms of oral mucositis.

In certain preferred embodiments, the TRPV3 antagonists is administeredto promote loss of or inhibit the growth of hair on a patient.

Still another aspect of the present invention relates to the use of aTRPV3 antagonist, e.g., a small molecule agent that inhibits inwardTRPV3-mediated current with an IC₅₀ of 1 micromolar or less, in themanufacture of a medicament to prevent, treat or alleviate symptoms of adisease, disorder or condition involving activation of TRPV3, or forwhich reduced TRPV3 activity can reduce the severity, in a patient.

Yet another aspect of the present invention relates to a pharmaceuticalpreparation comprising an agent that inhibits inward TRPV3-mediatedcurrent with an IC₅₀ of 1 micromolar or less; and a pharmaceuticallyacceptable excipient or solvent wherein the agent is provided in adosage form providing an amount effective to prevent, treat or alleviatesymptoms of a disease, disorder or condition involving activation ofTRPV3, or for which reduced TRPV3 activity can reduce the severity, in apatient. In certain preferred embodiments, the pharmaceuticalpreparation does not cause QT interval elongation in the patient.

In certain illustrative embodiments, the pharmaceutical preparationcomprises an agent that inhibits TRPV3-mediated current with an IC₅₀ ofat least one order of magnitude lower than its IC₅₀ for inhibition ofNaV 1.2 function, TRPV1 function, TRPV5 function, TRPV6 function,mitochondrial uniporter function and HERG function; and apharmaceutically acceptable excipient or solvent, wherein the agent isprovided in a dosage form providing an amount effective to prevent,treat or alleviate symptoms of a disease, disorder or conditioninvolving activation of TRPV3, or for which reduced TRPV3 acitivity canreduce the severity, in a patient, but which does not cause QT intervalelongation.

In another illustrative embodiment, the pharmaceutical preparationcomprises an agent that inhibits heat-induced TRPV3-mediated currentwith an IC₅₀ of 1 micromolar or less; and a pharmaceutically acceptableexcipient or solvent, wherein the agent is provided in a dosage formproviding an amount effective to prevent, treat or alleviate symptoms ofa disease, disorder or condition involving activation of TRPV3, or forwhich reduced TRPV3 activity can reduce the severity, in a patient, butwhich does not cause QT interval elongation.

One preferred preparation is a topical formulation for reducing TRPV3activity in skin or mucosa, comprising an agent that inhibits both 2-APB(2-aminoethyl diphenylborinate) and heat induced TRPV3-mediated currentwith an IC₅₀ of 1 micromolar or less.

Another preferred preparation is a removable patch or bandage,comprising: (i) a polymeric base; and (ii) an agent that inhibits both2-APB and heat induced TRPV3-mediated current with an IC₅₀ of 1micromolar or less.

Still another illustrative formultation is a skin exfoliant compositionfor topical application to an animal subject comprising a topicalvehicle; one or more skin exfoliant ingredients selected from the groupconsisting of carboxylic acids, keto acids, α-hydroxy acids, β-hydroxyacids, retinoids, peroxides, and organic alcohols, said one or more skinexfoliant ingredients contained in a total amount of at least about 12%by weight and capable of inducing skin irritation and effectingexfoliation of the skin of said subject; and an agent that inhibits both2-APB and heat induced TRPV3-mediated current with an IC₅₀ of 1micromolar or less, which agent is provided in an amount effective foranalgesic, anti-irritant and/or anti-inflammatory effects when appliedto skin.

Yet another embodiment is an antitussive composition for peroraladministration comprising an agent that inhibits both 2-APB and heatinduced TRPV3-mediated current with an IC₅₀ of 1 micromolar or less, andan orally-acceptable pharmaceutical carrier in the form of anaqueous-based liquid, or solid dissolvable in the mouth, selected fromthe group consisting of syrup, elixer, suspension, spray, lozenge,chewable lozenge, powder, and chewable tablet. Such antitussivecompositions can include one or more additional agents for treatingcough, allergy or asthma symptom selected from the group consisting of:antihistamines, 5-lipoxygenase inhibitors, leukotriene inhibitors, H3inhibitors, β-adrenergic receptor agonists, xanthine derivatives,α-adrenergic receptor agonists, mast cell stabilizers, expectorants,NK1, NK2 and NK3 tachykinin receptor antagonists, and GABA_(B) agonists.

Still another embodiment is a metered dose aerosol dispenser containingan aerosol pharmaceutical composition for pulmonary or nasal deliverycomprising an agent that inhibits both 2-APB and heat inducedTRPV3-mediated current with an IC₅₀ of 1 micromolar or less. Forinstance, it can be a metered dose inhaler, a dry powder inhaler or anair-jet nebulizer.

Still another embodiment is an eye ointment or eyedrops for ocularadministration. Such ocular compositions may be useful for the treatmentor alleviation of ocular pain including pain resulting from eye abrasionor post-surgical pain.

The present invention also relates to certain novel compounds, includingpurified preparations of those compounds. For instance, the inventionprovides compounds of Formula VII or a salt, solvate, hydrate, oxidativemetabolite or prodrug thereof:

wherein: X represents O, S, N(CN), CF₂, or C(CN)₂; Z represents O, S,N(CN), CF₂, or C(CN)₂; A represents N or C(R₂); G represents N or C(R₃);R₁ represents H or a substituent; R₂′ represents a substituent; R₂represents, independently for each occurrence, H or a substituent; R₃represents, independently for each occurrence, H or a substituent; and Drepresents O or N. In certain preferred embodiments: X represents S andD represents O; and/or R₁ is selected from H, lower alkyl, alkoxy,carboxyl, ester, amido, sulfonamido, heterocyclyl, cycloalkyl, hydroxyl,amino, acylamino, thioether, sulfonylamino, or R₁ taken together withthe carbon to which it is attached forms a carbonyl or thiocarbonyl; andR₂′, R₂ and R₃, independently for each occurrence, are selected from H,lower alkyl, alkoxy, carboxyl, ester, amido, sulfonamido, heterocyclyl,cycloalkyl, hydroxyl, amino, acylamino, thioether, sulfonylamino, nitro,halogen, CF₃, or cyano.

In certain embodiments, the compound can be represented by Formula VIIa:

wherein: R₁ represents H or a substituent; R₂ and R₃ are absent orrepresent one or more substituents on the ring to which they areattached; and R₂′ represents a substituent. In certain preferredembodiments, R₁ is selected from H, lower alkyl, alkoxy, carboxyl,ester, amido, sulfonamido, heterocyclyl, cycloalkyl, hydroxyl, amino,acylamino, thioether, sulfonylamino, or R₁ taken together with thecarbon to which it is attached forms a carbonyl or thiocarbonyl; R₂ andR₃ are absent or, independently for each occurrence, are selected fromlower alkyl, alkoxy, carboxyl, ester, amido, sulfonamido, heterocyclyl,cycloalkyl, hydroxyl, amino, acylamino, thioether, sulfonylamino, nitro,halogen, CF₃, or cyano; and R₂′ is selected from lower alkyl, alkoxy,carboxyl, ester, amido, sulfonamido, heterocyclyl, cycloalkyl, hydroxyl,amino, acylamino, thioether, sulfonylamino, halogen, CF₃, or cyano. Inmore preferred embodiments, each R₁ is independently selected from H,lower alkyl, or alkoxy, or R₁ taken together with the carbon to which itis attached forms a carbonyl. In certain embodiments, each R₂ is absentor is independently selected from lower alkyl, alkoxy, or halogen; R₂′is selected from lower alkyl, alkoxy, or halogen; and each R₃ is absentor is independently selected from lower alkyl, alkoxy, CF₃, nitro, orhalogen.

In certain preferred embodiments, R₁ and R₂ are absent; R₂′ representsCH₃; and R₃ represents one or more substituents positioned meta or parato the nitrogen of the ring system.

In certain embodiments, the compound can be represented by Formula VIIb:

wherein: R₁ represents H or a substituent; R₂ and R₃ are absent orrepresent one or more substituents on the ring to which they areattached; and R₂′ represents a substituent.

For instance, R₁ can be selected from H, lower alkyl, alkoxy, carboxyl,ester, amido, sulfonamido, heterocyclyl, cycloalkyl, hydroxyl, amino,acylamino, thioether, sulfonylamino, or R₁ taken together with thecarbon to which it is attached forms a carbonyl or thiocarbonyl; R₂ andR₃ are absent or, independently for each occurrence, are selected fromlower alkyl, alkoxy, carboxyl, ester, amido, sulfonamido, heterocyclyl,cycloalkyl, hydroxyl, amino, acylamino, thioether, sulfonylamino, nitro,halogen, CF₃, or cyano; and R₂′ is selected from lower alkyl, alkoxy,carboxyl, ester, amido, sulfonamido, heterocyclyl, cycloalkyl, hydroxyl,amino, acylamino, thioether, sulfonylamino, halogen, CF₃, or cyano.

In certain preferred embodiments, R₁ is independently selected from H,lower alkyl, or alkoxy, or R₁ taken together with the carbon to which itis attached forms a carbonyl.

In certain preferred embodiments, R₂ is absent or each R₂ isindependently selected from lower alkyl, alkoxy, or halogen.

In certain preferred embodiments, R₂′ is selected from lower alkyl,alkoxy, or halogen.

In certain preferred embodiments, R₃ is absent or each R₃ isindependently selected from lower alkyl, alkoxy, CF₃, nitro, or halogen.

In certain preferred embodiments, R₁, R₂, and R₃ are absent.

In certain embodiments, the TRPV3 antagonist can be represented byFormula VIII or a salt, solvate, hydrate, oxidative metabolite orprodrug thereof:

wherein: X represents O, S, N(CN), CF₂, or C(CN)₂; G represents N orC(R₃); R₁ and R₂, each independently, is absent or represents one ormore substituents on the ring to which it is attached; Ar′ represents anaryl or heteroaryl group; n represents 1, 2, or 3; R₃ represents,independently for each occurrence, H or a substituent; R₅ representsCH₂, O, NR₆; R₆ represents H or lower alkyl; and D represents O or N.

In certain embodiments, Ar′ represents phenyl, thiophene, imidazole,thiazole, furan, oxazole, tetrazole, thiadiazole, pyridine, pyrimadine,or triazole.

In certain embodiments, X represents S and D represents O.

In certain embodiments, R₁ is absent or, independently for eachoccurrence, is selected from lower alkyl, alkoxy, carboxyl, ester,amido, sulfonamido, heterocyclyl, cycloalkyl, hydroxyl, amino,acylamino, thioether, sulfonylamino, or R₁ taken together with thecarbon to which it is attached forms a carbonyl or thiocarbonyl; and R₂and R₃, independently for each occurrence, are selected from H, loweralkyl, alkoxy, carboxyl, ester, amido, sulfonamido, heterocyclyl,cycloalkyl, hydroxyl, amino, acylamino, thioether, sulfonylamino, nitro,halogen, CF₃, or cyano.

In certain embodiments, the TRPV3 antagonist can be represented byFormula VIIIa:

wherein: R₁, R₂ and R₃ are absent or represent one or more substituentson the ring to which they are attached.

In certain embodiments, R₁ is absent or, independently for eachoccurrence, is selected from lower alkyl, alkoxy, carboxyl, ester,amido, sulfonamido, heterocyclyl, cycloalkyl, hydroxyl, amino,acylamino, thioether, sulfonylamino, or R₁ taken together with thecarbon to which it is attached forms a carbonyl or thiocarbonyl; and R₂and R₃ are absent or, independently for each occurrence, are selectedfrom lower alkyl, alkoxy, carboxyl, ester, amido, sulfonamido,heterocyclyl, cycloalkyl, hydroxyl, amino, acylamino, thioether,sulfonylamino, nitro, halogen, CF₃, or cyano.

In certain embodiments, the TRPV3 antagonist is a compound of Formula IXor a salt, solvate, hydrate, oxidative metabolite or prodrug thereof:

wherein: A represents N or C(R₂); G represents N or C(R₃); R₁ representsH or a substituent; R₂ represents, independently for each occurrence, Hor a substituent; R₃ represents, independently for each occurrence, H ora substituent; R₃′ represents H, halogen, alkoxy, lower alkyl,perfluoroalkyl, aryl, heteroaryl, or CHR_(3a); R₃″ represents H,halogen, lower alkyl, aryl, heteroaryl, or CHR_(3a); R_(3a) representsamino, heterocyclyl, carboxyl, ester, aryl, or heteroaryl; and Drepresents O or N, provided that when R₃′ is H, R₃″ is not H or loweralkyl, and when R₃′ is halogen, R₃″ is not H.

In certain embodiments, R₁ is selected from H, lower alkyl, alkoxy,carboxyl, ester, amido, sulfonamido, heterocyclyl, cycloalkyl, hydroxyl,amino, acylamino, thioether, sulfonylamino, or R₁ taken together withthe carbon to which it is attached forms a carbonyl or thiocarbonyl; andR₂ and R₃, independently for each occurrence, are selected from H, loweralkyl, alkoxy, carboxyl, ester, amido, sulfonamido, heterocyclyl,cycloalkyl, hydroxyl, amino, acylamino, thioether, sulfonylamino,halogen, CF₃, or cyano.

In certain embodiments, the TRPV3 antagonist is a compound can berepresented by formula IXa

wherein R₁ represents H or a substituent; R₂ is absent or represents asubstituent; R₃ is absent or represents a substituent; R₃′ represents H,halogen, alkoxy, lower alkyl, perfluoroalkyl, aryl, heteroaryl, orCHR_(3a); R₃″ represents H, halogen, lower alkyl, aryl, heteroaryl, orCHR_(3a); R_(3a) represents amino, heterocyclyl, carboxyl, ester, aryl,or heteroaryl; and D represents O or N, provided that when R₃′ is H, R₃″is not H or lower alkyl, and when R₃′ is halogen, R₃″ is not H.

In certain embodiments, R₁ is selected from H, lower alkyl, alkoxy,carboxyl, ester, amido, sulfonamido, heterocyclyl, cycloalkyl, hydroxyl,amino, acylamino, thioether, sulfonylamino, or R₁ taken together withthe carbon to which it is attached forms a carbonyl or thiocarbonyl; andR₂ and R₃ are absent or, independently for each occurrence, are selectedfrom lower alkyl, alkoxy, carboxyl, ester, amido, sulfonamido,heterocyclyl, cycloalkyl, hydroxyl, amino, acylamino, thioether,sulfonylamino, halogen, CF₃, or cyano.

In certain embodiments, the TRPV3 antagonist is a compound of Formula Xor a salt, solvate, hydrate, oxidative metabolite or prodrug thereof:

wherein: X represents O, S, N(CN), CF₂, or C(CN)₂; Z represents O, S,N(CN), CF₂, or C(CN)₂; A represents N or C(R₂); G represents N or C(R₃);R₁ represents H or a substituent; R₂ represents, independently for eachoccurrence, H or a substituent; R₃ represents, independently for eachoccurrence, H or a substituent; and D represents O or N.

In certain embodiments, X represents S and D represents O.

In certain embodiments, R₁ is selected from H, lower alkyl, alkoxy,carboxyl, ester, amido, sulfonamido, heterocyclyl, cycloalkyl, hydroxyl,amino, acylamino, thioether, sulfonylamino, or R₁ taken together withthe carbon to which it is attached forms a carbonyl or thiocarbonyl; andR₂ and R₃, independently for each occurrence, are selected from H, loweralkyl, alkoxy, carboxyl, ester, amido, sulfonamido, heterocyclyl,cycloalkyl, hydroxyl, amino, acylamino, thioether, sulfonylamino, nitro,halogen, CF₃, or cyano.

In certain embodiments, the TRPV3 antagonist is a compound can berepresented by Formula Xa:

wherein: R₁ represents H or a substituent; and R₂ and R₃ are absent orrepresent one or more substituents on the ring to which they areattached.

In certain embodiments, R₁ is selected from H, lower alkyl, alkoxy,carboxyl, ester, amido, sulfonamido, heterocyclyl, cycloalkyl, hydroxyl,amino, acylamino, thioether, sulfonylamino, or R₁ taken together withthe carbon to which it is attached forms a carbonyl or thiocarbonyl; andR₂ and R₃ are absent or, independently for each occurrence, are selectedfrom lower alkyl, alkoxy, carboxyl, ester, amido, sulfonamido,heterocyclyl, cycloalkyl, hydroxyl, amino, acylamino, thioether,sulfonylamino, nitro, halogen, CF₃, or cyano.

In certain embodiments, R₁ is H; and R₂ and R₃ are absent.

In certain embodiments, the TRPV3 antagonist is a compound representedby Formula Xb:

wherein: R₁ represents H or a substituent; and R₂ and R₃ are absent orrepresent one or more substituents on the ring to which they areattached.

In certain embodiments, wherein R₁ is selected from H, lower alkyl,alkoxy, carboxyl, ester, amido, sulfonamido, heterocyclyl, cycloalkyl,hydroxyl, amino, acylamino, thioether, sulfonylamino, or R₁ takentogether with the carbon to which it is attached forms a carbonyl orthiocarbonyl; and R₂ and R₃ are absent or, independently for eachoccurrence, are selected from lower alkyl, alkoxy, carboxyl, ester,amido, sulfonamido, heterocyclyl, cycloalkyl, hydroxyl, amino,acylamino, thioether, sulfonylamino, nitro, halogen, CF₃, or cyano.

In certain embodiments, the TRPV3 antagonist is a compound of Formula XIor a salt, solvate, hydrate, oxidative metabolite or prodrug thereof:

wherein: X represents O, S, N(CN), CF₂, or C(CN)₂; Z represents O, S,N(CN), CF₂, or C(CN)₂; A represents N or C(R₂); G represents N or C(R₃);R₁ represents H or a substituent; R₂ represents, independently for eachoccurrence, H or a substituent; R₃ represents, independently for eachoccurrence, H or a substituent; R_(3b) represents a substituent; and Drepresents O or N.

In certain embodiments, X represents S and D represents O.

In certain embodiments, R₁ is selected from H, lower alkyl, alkoxy,carboxyl, ester, amido, sulfonamido, heterocyclyl, cycloalkyl, hydroxyl,amino, acylamino, thioether, sulfonylamino, or R₁ taken together withthe carbon to which it is attached forms a carbonyl or thiocarbonyl; andR₂ and R₃ are absent or, independently for each occurrence, are selectedfrom lower alkyl, alkoxy, carboxyl, ester, amido, sulfonamido,heterocyclyl, cycloalkyl, hydroxyl, amino, acylamino, thioether,sulfonylamino, nitro, halogen, CF₃, or cyano; and R_(3b) is selectedfrom lower alkyl, alkoxy, carboxyl, ester, amido, sulfonamido,heterocyclyl, cycloalkyl, hydroxyl, amino, acylamino, thioether,sulfonylamino, halogen, CF₃, or cyano.

In certain embodiments, the TRPV3 antagonist is a compound representedby Formula XIa:

wherein: R₁ represents H or a substituent; R₂ and R₃ are absent orrepresent one or more substituents on the ring to which they areattached; and R_(3b) represents a substituent.

In certain embodiments, R₁ is selected from H, lower alkyl, alkoxy,carboxyl, ester, amido, sulfonamido, heterocyclyl, cycloalkyl, hydroxyl,amino, acylamino, thioether, sulfonylamino, or R₁ taken together withthe carbon to which it is attached forms a carbonyl or thiocarbonyl; R₂and R₃ are absent or, independently for each occurrence, are selectedfrom lower alkyl, alkoxy, carboxyl, ester, amido, sulfonamido,heterocyclyl, cycloalkyl, hydroxyl, amino, acylamino, thioether,sulfonylamino, nitro, halogen, CF₃, or cyano; and R_(3b) is selectedfrom lower alkyl, alkoxy, carboxyl, ester, amido, sulfonamido,heterocyclyl, cycloalkyl, hydroxyl, amino, acylamino, thioether,sulfonylamino, halogen, CF₃, or cyano.

In certain embodiments, R₁ is H; R₂ and R₃ are absent; and R_(3b) is ahalogen.

In certain embodiments, the TRPV3 antagonist is a compound can berepresented by Formula XIb:

wherein: R₁ represents H or a substituent; R₂ and R₃ are absent orrepresent one or more substituents on the ring to which they areattached; and R_(3b) represents a substituent.

In certain embodiments, R₁ is selected from H, lower alkyl, alkoxy,carboxyl, ester, amido, sulfonamido, heterocyclyl, cycloalkyl, hydroxyl,amino, acylamino, thioether, sulfonylamino, or R₁ taken together withthe carbon to which it is attached forms a carbonyl or thiocarbonyl; R₂and R₃ are absent or, independently for each occurrence, are selectedfrom lower alkyl, alkoxy, carboxyl, ester, amido, sulfonamido,heterocyclyl, cycloalkyl, hydroxyl, amino, acylamino, thioether,sulfonylamino, nitro, halogen, CF₃, or cyano; and R_(3b) is selectedfrom lower alkyl, alkoxy, carboxyl, ester, amido, sulfonamido,heterocyclyl, cycloalkyl, hydroxyl, amino, acylamino, thioether,sulfonylamino, halogen, CF₃, or cyano.

DETAILED DESCRIPTION OF THE DRAWINGS

FIG. 1 provides a schematic representation of experiments performedusing a thermal injury model.

FIG. 2 summarizes experiments showing the effectiveness of a TRPV3inhibitor in diminishing pain in the thermal injury model. The reductionin pain is dose dependent.

FIG. 3 shows that the efficacy of a TRPV3 inhibitor in diminishing painin the thermal injury model is specific, and that a TRPV3 inhibitordiminishes pain without affecting the uninjured paw. This suggests thatTRPV3 inhibitors are specific for the facilitated state and arenon-narcotic.

FIG. 4 shows that, in contrast to a TRPV3 inhibitor, morphine reducespain but has a narcotic effect. Note the thermal escape latency aftertreatment with morphine is elevated above the baseline.

FIG. 5 provides a schematic representation of experiments performedusing an automated formalin model.

FIG. 6 summarizes experiments showing the effectiveness of a TRPV3inhibitor in diminishing pain in the formalin model. The TRPV3 inhibitorreduces pain during the second phase (also known as the Late Phase orthe facilitated phase) following formalin injection. There is also atrend towards a reduction in the first phase (also known as the EarlyPhase).

FIG. 7 provides a schematic representation of experiments performedusing the carrageenan model of acute inflammatory pain.

FIG. 8 summarizes experiments showing that a TRPV3 inhibitor reducesthermal hyperalgesia in the carrageenan model of acute inflammatorypain. The effectiveness of the TRPV3 inhibitor is similar to that of aTRPV1 inhibitor.

FIG. 9 provides a schematic representation of experiments performedusing the Complete Freund's Adjuvant (CFA) model of inflammatory pain.

FIG. 10 summarizes experiments showing that a TRPV3 inhibitor reducesinflammatory pain in the CFA model. The effectiveness of the TRPV3inhibitor is comparable or superior to that of diclofenac (anon-steroidal anti-inflammatory), as well as a TRPV1 inhibitor.

FIG. 11 summarizes experiments showing the responses for individualrats. TRPV3 inhibitor restores pain sensitivity to baseline afterinjection of CFA.

FIG. 12 summarizes experiments performed using a second TRPV3 inhibitor.This second TRPV3 inhibitor also reduces pain in the CFA model.Furthermore, the effectiveness of the TRPV3 inhibitor in reducing painin the CFA model is similar to that of the non-steroidalanti-inflammatory, diclofenac.

FIG. 13 a-13 b summarizes experiments measuring plasma levels of apotent TRPV3 inhibitor described in the example below. The TRPV3inhibitor was administered intravenously (a) or orally in a suspension(b).

FIG. 14 shows the SAR for various compounds of Formula I versus TRPV3.

FIG. 15 shows the SAR for various compounds of Formula IV versus TRPV3and in some instances versus other channels.

FIG. 16 shows the SAR for various compounds of Formula III versus TRPV3and in some instances versus other channels.

FIG. 17 shows the SAR for various compounds of Formula II versus TRPV3and in some instances versus other channels.

FIG. 18 shows the SAR for various compounds versus TRPV3.

DETAILED DESCRIPTION OF THE INVENTION

Cellular homeostasis is a result of the summation of regulatory systemsinvolved in, amongst other things, the regulation of ion flux andmembrane potential. Cellular homeostasis is achieved, at least in part,by movement of ions into and out of cells across the plasma membrane andwithin cells by movement of ions across membranes of intracellularorganelles including, for example, the endoplasmic reticulum,sarcoplasmic reticulum, mitochondria and endocytic organelles includingendosomes and lysosomes.

Movement of ions across cellular membranes is carried out by specializedproteins. TRP channels are one large family of non-selective cationchannels that function to help regulate ion flux and membrane potential.TRP channels are subdivided into 6 sub-families including the TRPV(vanilloid receptor) family. TRPV3 is a member of the TRPV class of TRPchannels.

Non-selective cation channels such as TRPV3 modulate the flux of calciumand sodium ions across cellular membranes. Sodium and calcium influxleads to a depolarization of the cell. This increases the probabilitythat voltage-gated ion channels will reach the threshold required foractivation. As a result, activation of non-selective cation channels canincrease electrical excitability and increase the frequency ofvoltage-dependent events. Voltage dependent events include, but are notlimited to, neuronal action potentials, cardiac action potentials,smooth muscle contraction, cardiac muscle contraction, and skeletalmuscle contraction.

TRPV3 is also highly expressed in skin. In a keratinocyte cell line,stimulation of TRPV3 leads to release of inflammatory mediatorsincluding interleukin-1. Thus TRPV3 may also play an important role inregulating inflammation and pain that results from the release ofinflammatory stimuli (Xu et al., 2006)

TRPV3 proteins are thermosensitive channels expressed in skin cells(see, e.g., Peier et al. (2002) Science 296:2046-2049) and dorsal rootganglion, trigeminal ganglion, spinal cord and brain (see, e.g., Xu etal. (2002) Nature 418:181-185; Smith et al. (2002) Nature 418:186-188).Particular TRPV3 proteins that may be used in screening assays, asdescribed herein, to identify compounds that modulate a function ofTRPV3 include, but are not limited to human TRPV3, mouse TRPV3, andDrosophila TRPV3. U.S. Patent Application Publication 2004/0009537 (the“'537 publication”) disclosed sequences corresponding to human, mouse,and Drosophila TRPV3. For example, SEQ ID NOs 106 and 107 of the '537publication correspond to the human nucleic acid and amino acidsequences, respectively. SEQ ID Nos 108 and 109 of the '537 publicationcorrespond to the mouse nucleic acid and amino acid sequences,respectively. The Drosophila protein is approximately 25% identical and41% homologous to the human protein over approximately 49% of the lengthof the protein, and approximately 26% identical and 42% homologous tothe mouse protein over approximately 49% of the length of the protein.

Other exemplary human TRPV3 nucleic acid and amino acid sequences aredisclosed in GenBank at the following accession numbers: gi:21912412(accession no. AJ487035); gi:21435923 (accession no. AF514998);gi:22651775 (accession no. AY118268); gi:85397600 (accession no.BC104868); and gi:22651773 (accession no. AY118267). The TRPV3 sequencesand disclosures provided at these accession numbers are herebyincorporated by reference in their entirety. Compounds that inhibit oneor more functions or activities of TRPV3, according to the presentinvention, inhibit one or more functions of any of the TRPV3 proteinsprovided herein. Furthermore, compounds that inhibit one or morefunctions or activities of TRPV3, according to the present invention,inhibit one or more functions of a TRPV3 protein encoded by a nucleicacid sequence that hybridizes under stringent conditions, including awash step of 0.2×SSC at 65° C., to a TRPV3 nucleic acid sequenceprovided herein.

TRPV3 is expressed in a pattern consistent with a role in, among otherthings, pain. TRPV3 is expressed in tissues containing pain-sensingneurons (nociceptors). Nociceptors mediate responsiveness to force,heat, cold, chemicals, and inflammation. In addition, skin whichexpresses high levels of TRPV3 plays a significant role in pain.Additional evidence implicating TRPV3 in, among other processes, painincludes the TRPV3 knock-out mouse which displays abnormal responses topainful stimuli. Further evidence indicates that TRPV3 expressionincreases in the skin cells of breast cancer patients who reportsignificant pain (Gopinath et al., 2005).

Accordingly, modulating the function of TRPV3 proteins provides a meansof modulating calcium homeostasis, sodium homeostasis, membranepolarization, and/or intracellular calcium levels, and compounds thatcan modulate TRPV3 function are useful in many aspects, including, butnot limited to, maintaining calcium homeostasis, modulatingintracellular calcium levels, modulating membrane polarization, andtreating or preventing diseases, disorders, or conditions associatedwith calcium and/or sodium homeostasis or dyshomeostasis. In oneembodiment, compounds that modulate TRPV3 function can be used in thetreatment of diseases, injuries, disorders, or conditions caused orexacerbated, in whole or in part, by regulation or misregulation ofTRPV3 activity. In one embodiment, compounds that inhibit a TRPV3function can be used in the treatment of diseases, injuries, disorders,or conditions caused or exacerbated, in whole or in part, by regulationor misregulation of TRPV3 activity. In still another embodiment,compounds that inhibit a TRPV3 function can be used in the treatment ofpain.

In certain embodiments, the TRPV3 antagonist is “small molecule”, e.g.,an organic molecule having a molecular weight of 2000 amu or less.Exemplary TRPV3 antagonists include a compound of Formula I or a salt,solvate, hydrate, oxidative metabolite or prodrug thereof:

wherein: Ar represents an aryl or heteroaryl group; Y represents Ph,OArl, SArl, or N(U)(Arl); R represents H or a lower alkyl; X representsCH₂, O, S, NR₁₀, CF₂, or C(CN)₂; W represents O, S, or NR′; n is 1, orwhen X is CH₂, n is 1 or 2; Arl represents an aralkyl, heteroaralkyl,aryl, or heteroaryl group, which may be monocyclic or bicyclic; Phrepresents a substituted or unsubstituted phenyl ring, optionally fusedto a substituted or unsubstituted 5- to 7-membered cycloalkyl,heterocyclyl, aryl, or heteroaryl ring; R′, independently for eachoccurrence, represents H or a lower alkyl; U represents hydrogen orlower alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl,heteroaryl, cycloalkylalkyl, heterocyclylalkyl, aralkyl, orheteroaralkyl, or, U, N and R′ taken together form a 5- to 7-memberedheterocyclic ring, or, U, Arl and N taken together form a ring fused tothe aryl or heteroaryl ring of Arl, thereby forming a bicyclicstructure; R₁₀ represents H, lower alkyl, or cyano; and wherein saidcompound inhibits TRPV3 with an with an IC₅₀ of 10 micromolar or less.

In certain embodiments, R is H, W is O or S, and X is O or S.

In certain embodiments, X is CH₂.

In certain preferred embodiments, the TRPV3 antagonist is represented inFormula Ia, or is a salt, solvate, hydrate, oxidative metabolite orprodrug thereof:

wherein: Ar represents an aryl or heteroaryl group; Y represents Ph,OArl, SArl, or N(U)(Arl); R represents H or a lower alkyl; X representsO, S, NR₁₀, CF₂, or C(CN)₂; W represents O, S, or NR′; Arl represents anaralkyl, heteroaralkyl, aryl, or heteroaryl group, which may bemonocyclic or bicyclic; Ph represents a substituted or unsubstitutedphenyl ring, optionally fused to a substituted or unsubstituted 5- to7-membered cycloalkyl, heterocyclyl, aryl, or heteroaryl ring; R′,independently for each occurrence, represents H or a lower alkyl; Urepresents hydrogen or lower alkyl, alkenyl, alkynyl, cycloalkyl,heterocyclyl, aryl, heteroaryl, cycloalkylalkyl, heterocyclylalkyl,aralkyl, or heteroaralkyl, or, U, N and R′ taken together form a 5- to7-membered heterocyclic ring, or, U, Arl and N taken together form aring fused to the aryl or heteroaryl ring of Arl, thereby forming abicyclic structure; and R₁₀ represents H, lower alkyl, or cyano.

Exemplary compounds of Formula I and Ia include:

Other exemplary compounds of Formula I are shown in FIG. 14. FIG. 14shows the SAR for various compounds of Formula I versus TRPV3.

In certain embodiments of Formulas I and Ia, Y is N(U)(Arl), and in suchembodiments, Y can represent, for example,

wherein: R₁ and R₂, each independently, is absent or represents one ormore substituents on the ring to which it is attached; Ar′ represents anaryl or heteroaryl group; R₅ represents CH₂, O, NR₆; R₆ represents H orlower alkyl; and n represents 1, 2, or 3. In certain embodiments of theabove-referenced compounds, one or more of the following conditions maybe true:

-   -   Ar′ represents phenyl, thiophene, imidazole, thiazole, furan,        oxazole, tetrazole, thiadiazole, pyridine, pyrimadine, or        triazole;    -   each R₁ is independently selected from H, lower alkyl, alkoxy,        carboxyl, ester, amido, sulfonamido, heterocyclyl, cycloalkyl,        hydroxyl, amino, acylamino, thioether, sulfonylamino, or R₁        taken together with the carbon to which it is attached forms a        carbonyl or thiocarbonyl;    -   each R₂ is independently selected from H, lower alkyl, alkoxy,        carboxyl, ester amido, sulfonamido, heterocyclyl, cycloalkyl,        hydroxyl, amino, acylamino, thioether, sulfonylamino, halogen,        CF₃, or cyano;

In certain preferred embodiments of Formulas I and Ia, Y represents

wherein R₁ and R₂ each, independently, are absent or represent one ormore substituents on the ring to which they are attached. Even morepreferably, independently for each occurrence, R₁ is selected from H,lower alkyl, alkoxy, carboxyl, ester, amido, sulfonamido, heterocyclyl,cycloalkyl, hydroxyl, amino, acylamino, thioether, sulfonylamino, or R1taken together with the carbon to which it is attached forms a carbonylor thiocarbonyl; and independently for each occurrence, R₂ is selectedfrom H, lower alkyl, alkoxy, carboxyl, ester, amido, sulfonamido,heterocyclyl, cycloalkyl, hydroxyl, amino, acylamino, thioether,sulfonylamino, halogen, CF₃, or cyano.

For further illustration, Y can represent

wherein R₁ represents H, lower alkyl, alkoxy, carboxyl, ester, amido,sulfonamido, heterocyclyl, cycloalkyl, hydroxyl, amino, acylamino,thioether, sulfonylamino, or R₁ taken together with the carbon to whichit is attached forms a carbonyl or thiocarbonyl; and R₂ represents H,lower alkyl, alkoxy, carboxyl, ester, amido, sulfonamido, heterocyclyl,cycloalkyl, hydroxyl, amino, acylamino, thioether, sulfonylamino,halogen, CF₃, or cyano. In certain preferred embodiments, R₁ representsH, lower alkyl, alkoxy, or R₁ taken together with the carbon to which itis attached forms a carbonyl, and R₂ represents H, lower alkyl, alkoxyor halogen.

In certain preferred embodiments of Formulas I and Ia, Ar represents

wherein: Cy represents an aryl or heteroaryl ring; Z represents O, S,NR₁₁, CF₂, or C(CN)₂; R₃ is absent or represents one or moresubstituents on the ring to which it is attached; R₁₁ represents H,lower alkyl, or cyano; A represents N or C(R_(2a)); and R_(2a)represents H, lower alkyl, alkoxy, or halogen.

For example, Cy can represent a phenyl, thiophene, imidazole, thiazole,furan, oxazole, tetrazole, thiadiazole, pyridine, pyrimadine, ortriazole, and R₃ can be absent, or independently for each occurrence,selected from lower alkyl, alkoxy, carboxyl, ester, amido, sulfonamido,heterocyclyl, cycloalkyl, hydroxyl, amino, acylamino, thioether,sulfonylamino, nitro, halogen, CF₃, or cyano. In certain preferredembodiments, A represents N, and Z represents O or S.

To further illustrate, Ar can represent

wherein: Z represents O, S, NR₁₁, CF₂, NR, or C(CN)₂; A represents N orC(R_(2a)); R₃ is absent or represents one or more substituents on thering to which it is attached; R₁₁ represents H, lower alkyl, or cyano;and R_(2a) represents H, lower alkyl, alkoxy, or halogen. In certainpreferred embodiments: Z represents O or S; and R₃ is absent or each R₃is independently selected from lower alkyl, alkoxy, carboxyl, ester,amido, sulfonamido, heterocyclyl, cycloalkyl, hydroxyl, amino,acylamino, thioether, sulfonylamino, nitro, halogen, CF₃, or cyano;

In certain preferred embodiments, Ar can represent

wherein: Z represents O, S, NR₁₁, CF₂, or C(CN)₂; A represents N orC(R_(2a)); R₃ independently for each occurrence represents H, loweralkyl, alkoxy, carboxyl, ester, amido, sulfonamido, heterocyclyl,cycloalkyl, hydroxyl, amino, acylamino, thioether, sulfonylamino,halogen, CF₃, or cyano; R₁₁ represents H, lower alkyl, or cyano; andR_(2a) represents H, lower alkyl, alkoxy, or halogen. Preferrably Zrepresents O or S, and R₃ independently for each occurrence representsH, lower alkyl, alkoxy, CF₃, nitro, or halogen.

In certain other preferred embodiments of Formulas I and Ia, Arrepresents

wherein: G′ represents N or C(R₅); R₃ is absent or represents one ormore substituents on Cy; R₅ represents H or a substituent; R₄ representsH or a substituent; and Cy represents an aryl or heteroaryl ring. Incertain preferred embodiments, Cy can be a phenyl, thiophene, imidazole,thiazole, furan, oxazole, tetrazole, thiadiazole, pyridine, pyrimadine,or triazole. In certain embodiments, R₃ is absent, or independently foreach occurrence, is selected from lower alkyl, alkoxy, carboxyl, ester,amido, sulfonamido, heterocyclyl, cycloalkyl, hydroxyl, amino,acylamino, thioether, sulfonylamino, nitro, halogen, CF₃, or cyano; andR₄ and R₅, independently for each occurrence, are selected from H, loweralkyl, alkoxy, carboxyl, ester, amido, sulfonamido, heterocyclyl,cycloalkyl, hydroxyl, amino, acylamino, thioether, sulfonylamino,halogen, CF₃, or cyano.

In still other embodiments, the TRPV3 antagonist is a compound ofFormula V or a salt, solvate, hydrate, oxidative metabolite or prodrugthereof:

wherein: X represents O, S, NR₁₀, CF₂, or C(CN)₂; Z represents O, S,NR₁₁, CF₂, or C(CN)₂; R₁₀ represents H, lower alkyl, or cyano; R₁₁represents H, lower alkyl, or cyano; A represents N or C(R₂); Grepresents N or C(R₃); R₁ is absent or represents one or moresubstituents on the ring to which it is attached; R₂ represents,independently for each occurrence, H or a substituent; R₃ represents,independently for each occurrence, H or a substituent; and D representsO or N.

In certain preferred embodiments of formula V: X represents S and Drepresents O; R₁ is absent, or independently for each occurrence, isselected from lower alkyl, alkoxy, carboxyl, ester, amido, sulfonamido,heterocyclyl, cycloalkyl, hydroxyl, amino, acylamino, thioether,sulfonylamino, or R₁ taken together with the carbon to which it isattached forms a carbonyl or thiocarbonyl; and R₂ and R₃, independentlyfor each occurrence, are selected from H, lower alkyl, alkoxy, carboxyl,ester, amido, sulfonamido, heterocyclyl, cycloalkyl, hydroxyl, amino,acylamino, thioether, sulfonylamino, nitro, halogen, CF₃, or cyano.

To further illustrate, the TRPV3 antagonist can be represented byFormula Va:

wherein: R₁, R₂ and R₃ are absent or represent one or more substituentson the ring to which they are attached. For instance, R₁ is absent, orindependently for each occurrence, is selected from lower alkyl, alkoxy,carboxyl, ester, amido, sulfonamido, heterocyclyl, cycloalkyl, hydroxyl,amino, acylamino, thioether, sulfonylamino, or R₁ taken together withthe carbon to which it is attached forms a carbonyl or thiocarbonyl; andR₂ and R₃ are absent, or independently for each occurrence, are selectedfrom lower alkyl, alkoxy, carboxyl, ester, amido, sulfonamido,heterocyclyl, cycloalkyl, hydroxyl, amino, acylamino, thioether,sulfonylamino, nitro, halogen, CF₃, or cyano. In certain preferredembodiments, R₁ is absent, or independently for each occurrence, isselected from lower alkyl, or alkoxy, or R₁ taken together with thecarbon to which it is attached forms a carbonyl; R₂ is absent, orindependently for each occurrence, is selected from lower alkyl, alkoxy,or halogen; and R₃ is absent, or independently for each occurrence, isselected from lower alkyl, alkoxy, CF₃, nitro, or halogen. In even morepreferred embodiments, R₁ represents H; R₂ represents H or CH₃ and ispositioned ortho to the N of the ring system; and R₃ represents one ormore substituents positioned meta or para to the nitrogen of the ringsystem.

In still another illustration, the TRPV3 antagonist can be representedby Formula Vb:

wherein R₁, R₂ and R₃ are absent or represent one or more substituentson the ring to which they are attached. For instance, R₁ is absent, orindependently for each occurrence, is selected from lower alkyl, alkoxy,carboxyl, ester, amido, sulfonamido, heterocyclyl, cycloalkyl, hydroxyl,amino, acylamino, thioether, sulfonylamino, or R₁ taken together withthe carbon to which it is attached forms a carbonyl or thiocarbonyl; andR₂ and R₃ are absent, or independently for each occurrence, are selectedfrom lower alkyl, alkoxy, carboxyl, ester, amido, sulfonamido,heterocyclyl, cycloalkyl, hydroxyl, amino, acylamino, thioether,sulfonylamino, nitro, halogen, CF₃, or cyano. In certain preferredembodiments, R₁ is absent, or independently for each occurrence, isselected from lower alkyl, or alkoxy, or R₁ taken together with thecarbon to which it is attached forms a carbonyl; R₂ is absent, orindependently for each occurrence, is selected from lower alkyl, alkoxy,or halogen; and R₃ is absent, or independently for each occurrence, isselected from lower alkyl, alkoxy, CF₃, nitro, or halogen. In certainembodiments, R₁, R₂, and R₃ are absent.

In still other embodiments, the TRPV3 antagonist is a compound ofFormula VI or a salt, solvate, hydrate, oxidative metabolite or prodrugthereof:

wherein: X represents O, S, NR₁₀, CF₂, or C(CN)₂; R₁₀ represents H,lower alkyl, or cyano; A represents N or C(R₂); G represents N or C(R₃);R₁ is absent or represents one or more substituents on the ring to whichit is attached; R₂ represents, independently for each occurrence, H or asubstituent; R₃ represents, independently for each occurrence, H or asubstituent; R₄ represents H or a substituent; D represents O or N; andwherein said compound inhibits TRPV3 with an with an IC₅₀ of 10micromolar or less.

In certain preferred embodiments, X represents S and D represents O.

In certain embodiments, the compound can be represented by Formula VIa:

wherein: R₁, R₂ and R₃ are absent or represent one or more substituentson the ring to which they are attached; and R₄ represents H or asubstituent. In certain preferred embodiments, R₁ is absent, orindependently for each occurrence, is selected from lower alkyl, alkoxy,carboxyl, ester, amido, sulfonamido, heterocyclyl, cycloalkyl, hydroxyl,amino, acylamino, thioether, sulfonylamino, or R₁ taken together withthe carbon to which it is attached forms a carbonyl or thiocarbonyl; andR₂, R₃ and R₄ are absent, or independently for each occurrence, isselected from H, lower alkyl, alkoxy, carboxyl, ester, amido,sulfonamido, heterocyclyl, cycloalkyl, hydroxyl, amino, acylamino,thioether, sulfonylamino, nitro, halogen, CF₃, or cyano. In preferredembodiments, the R₁ is absent, or independently for each occurrence, isselected from lower alkyl, or alkoxy, or R₁ taken together with thecarbon to which it is attached forms a carbonyl; R₂ is absent, orindependently for each occurrence, is selected from lower alkyl, alkoxy,or halogen; R₃ absent, or independently for each occurrence, is selectedfrom H, lower alkyl, alkoxy, CF₃, nitro, or halogen; and R₄ is selectedfrom H, lower alkyl, or cyano. In other preferred embodiments, R₁ and R₂are absent; R₃ represents methoxy positioned para to the N of the ringsystem; and R₄ represents cyano.

In certain embodiments, the compound can be represented by Formula VIb:

wherein: R₁, R₂ and R₃ are absent or represent one or more substituentson the ring to which they are attached; and R₄ represents H or asubstituent. For instance, R₁ is absent, or independently for eachoccurrence, is selected from lower alkyl, alkoxy, carboxyl, ester,amido, sulfonamido, heterocyclyl, cycloalkyl, hydroxyl, amino,acylamino, thioether, sulfonylamino, or R₁ taken together with thecarbon to which it is attached forms a carbonyl or thiocarbonyl; and R₂,R₃ and R₄ are absent, or independently for each occurrence, is selectedfrom lower alkyl, alkoxy, carboxyl, ester, amido, sulfonamido,heterocyclyl, cycloalkyl, hydroxyl, amino, acylamino, thioether,sulfonylamino, nitro, halogen, CF₃, or cyano. In certain preferredembodiments, R₁ is absent, or independently for each occurrence, isselected from lower alkyl, alkoxy, or halogen or R₁ taken together withthe carbon to which it is attached forms a carbonyl; R₂ is absent, orindependently for each occurrence, is selected from lower alkyl, alkoxy,or halogen; R₃ is absent, or independently for each occurrence, isselected from lower alkyl, alkoxy, CF₃, nitro, or halogen; and R₄ isselected from H, lower alkyl, or cyano.

In still other illustrative embodiments, the compound can be representedby Formula II or a salt, solvate, hydrate, oxidative metabolite orprodrug thereof:

wherein: W represents O or S; X′ represents NR₁₂; R₁₂ represents H orlower alkyl; Ar and Ar′ independently represent substituted orunsubstituted aryl or heteroaryl rings; and wherein said compoundinhibits TRPV3 with an with an IC₅₀ of 10 micromolar or less. Forinstance, Ar can represents phenyl, as can Ar′.

Examples of compounds within the above formula include:

FIG. 17 shows the SAR for various compounds of Formula II versus TRPV3and in some instances versus other channels.

In yet another illustrative embodiments, the compound can be representedby Formula III or a salt, solvate, hydrate, oxidative metabolite orprodrug thereof:

wherein: Ar represents a substituted or unsubstituted aryl or heteroarylring; Q represents Q, S, NR, or CH₂; R, independently for eachoccurrence, represents H or lower alkyl; R′ represents H or asubstituted or unsubstituted lower alkyl, alkenyl, alkynyl, cycloalkyl,heterocyclyl, aryl, heteroaryl, cycloalkylalkyl, heterocyclylalkyl,aralkyl, or heteroaralkyl; R″, independently for each occurrence,represents H or lower alkyl; V is absent or represents C═O, C═S, or SO₂;W represents a hydrogen-bond acceptor, such as ═O, ═S, amino, hydroxyl,or ether; E represents H, a substituted or unsubstituted lower alkyl,alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl,cycloalkylalkyl, heterocyclylalkyl, aralkyl, or heteroaralkyl, orhalogen, carbonyl (e.g., ester, carboxyl, or formyl), thiocarbonyl(e.g., thioester, thiocarboxylate, or thioformate), ketone, aldehyde,amino, acylamino, amido, amidino, cyano, nitro, azido, sulfonyl,sulfoxido, sulfate, sulfonate, sulfamoyl, sulfonamido, or phosphoryl; xrepresents 0 or 1 wherein when x is 0, the carbons bearing W and V aredirectly linked by a single or double bond; and wherein said compoundinhibits TRPV3 with an with an IC₅₀ of 10 micromolar or less.

In certain embodiments, the compounds of Formula III are characterizedby one or more of the following: Q represents S or CH₂; R represents H;R′ represents a substituted or unsubstituted aryl or heteroaryl ring; R″represents lower alkyl; and/or E represents CO₂Me or CO₂Et.

Exemplary compounds of Formula III include:

FIG. 16 shows the SAR for various compounds of Formula III versus TRPV3and in some instances versus other channels.

In yet another illustrative embodiments, the compound can be representedby Formula IIIA or a salt, solvate, hydrate, oxidative metabolite orprodrug thereof:

wherein: R, independently for each occurrence, represents H or loweralkyl, preferably H; R′ represents substituted or unsubstituted alkyl,alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl,cycloalkylalkyl, heterocyclylalkyl, aralkyl, or heteroaralkyl,preferably substituted or unsubstituted aryl or heteroaryl; R″,independently for each occurrence, represents H or lower alkyl,preferably lower alkyl, such as methyl; W represents O or S; Erepresents H, a substituted or unsubstituted alkyl, alkenyl, alkynyl,cycloalkyl, heterocyclyl, aryl, heteroaryl, cycloalkylalkyl,heterocyclylalkyl, aralkyl, or heteroaralkyl, or halogen, carbonyl(e.g., ester, carboxyl, or formyl), thiocarbonyl (e.g., thioester,thiocarboxylate, or thioformate), ketone, aldehyde, amino, acylamino,amido, amidino, cyano, nitro, azido, sulfonyl, sulfoxido, sulfate,sulfonate, sulfamoyl, sulfonamido, or phosphoryl, preferably acyl, acyano or nitro substituent, or a carboxylic acid or ester or amidederivative thereof, even more preferably a carboxylic acid or ester oramide derivative thereof, such as CO₂Me or CO₂Et; and Ar represents asubstituted or unsubstituted aryl or heteroaryl ring.

Additional illustrative embodiments of TRPV3 antagonists includecompounds can be represented by Formula IV or a salt, solvate, hydrate,oxidative metabolite or prodrug thereof:

wherein: Ar and Ar′ independently represent substituted or unsubstitutedaryl or heteroaryl rings; R, independently for each occurrence,represents H or lower alkyl; X, independently for each occurrence,represents O, S, NR, CF₂, or CR₂; M, independently for each occurrence,represents a substituted or unsubstituted methylene group; and whereinsaid compound inhibits TRPV3 with an with an IC₅₀ of 10 micromolar orless.

In certain embodiments, the compounds of Formula IV are characterized byone or more of the following: Ar and Ar′ represent phenyl or pyridylrings; R represents H; X represents O or S; M represents CH₂.

Examples of compounds within the above formula include:

FIG. 15 shows the SAR for various compounds of Formula IV versus TRPV3and in some instances versus other channels.

Additional illustrative embodiments of TRPV3 antagonists whichapplicants also observed to be TRPV3 inhibitors, include compounds whichcan be represented by the following structures or a salt, solvate,hydrate, oxidative metabolite or prodrug thereof:

FIG. 18 shows the SAR for various compounds versus TRPV3.

The present invention also relates to certain novel compounds, includingpurified preparations of those compounds. For instance, the inventionprovides compounds of Formula VII or a salt, solvate, hydrate, oxidativemetabolite or prodrug thereof:

wherein: X represents O, S, N(CN), CF₂, or C(CN)₂; Z represents O, S,N(CN), CF₂, or C(CN)₂; A represents N or C(R₂); G represents N or C(R₃);R₁ represents H or a substituent; R₂′ represents a substituent; R₂represents, independently for each occurrence, H or a substituent; R₃represents, independently for each occurrence, H or a substituent; and Drepresents O or N. In certain preferred embodiments: X represents S andD represents O; and/or R₁ is selected from H, lower alkyl, alkoxy,carboxyl, ester, amido, sulfonamido, heterocyclyl, cycloalkyl, hydroxyl,amino, acylamino, thioether, sulfonylamino, or R₁ taken together withthe carbon to which it is attached forms a carbonyl or thiocarbonyl; andR₂′, R₂ and R₃, independently for each occurrence, are selected from H,lower alkyl, alkoxy, carboxyl, ester, amido, sulfonamido, heterocyclyl,cycloalkyl, hydroxyl, amino, acylamino, thioether, sulfonylamino, nitro,halogen, CF₃, or cyano.

In certain embodiments, the compound can be represented by Formula VIIa:

wherein: R₁ represents H or a substituent; R₂ and R₃ are absent orrepresent one or more substituents on the ring to which they areattached; and R₂′ represents a substituent. In certain preferredembodiments, R₁ is selected from H, lower alkyl, alkoxy, carboxyl,ester, amido, sulfonamido, heterocyclyl, cycloalkyl, hydroxyl, amino,acylamino, thioether, sulfonylamino, or R₁ taken together with thecarbon to which it is attached forms a carbonyl or thiocarbonyl; R₂ andR₃ are absent or, independently for each occurrence, are selected fromlower alkyl, alkoxy, carboxyl, ester, amido, sulfonamido, heterocyclyl,cycloalkyl, hydroxyl, amino, acylamino, thioether, sulfonylamino, nitro,halogen, CF₃, or cyano; and R₂′ is selected from lower alkyl, alkoxy,carboxyl, ester, amido, sulfonamido, heterocyclyl, cycloalkyl, hydroxyl,amino, acylamino, thioether, sulfonylamino, halogen, CF₃, or cyano. Inmore preferred embodiments, each R₁ is independently selected from H,lower alkyl, or alkoxy, or R₁ taken together with the carbon to which itis attached forms a carbonyl. In certain embodiments, each R₂ is absentor is independently selected from lower alkyl, alkoxy, or halogen; R₂′is selected from lower alkyl, alkoxy, or halogen; and each R₃ is absentor is independently selected from lower alkyl, alkoxy, CF₃, nitro, orhalogen.

In certain preferred embodiments, R₁ and R₂ are absent; R₂′ representsCH₃; and R₃ represents one or more substituents positioned meta or parato the nitrogen of the ring system.

In certain embodiments, the compound can be represented by Formula VIIb:

wherein: R₁ represents H or a substituent; R₂ and R₃ are absent orrepresent one or more substituents on the ring to which they areattached; and R₂′ represents a substituent.

For instance, R₁ can be selected from H, lower alkyl, alkoxy, carboxyl,ester, amido, sulfonamido, heterocyclyl, cycloalkyl, hydroxyl, amino,acylamino, thioether, sulfonylamino, or R₁ taken together with thecarbon to which it is attached forms a carbonyl or thiocarbonyl; R₂ andR₃ are absent or, independently for each occurrence, are selected fromlower alkyl, alkoxy, carboxyl, ester, amido, sulfonamido, heterocyclyl,cycloalkyl, hydroxyl, amino, acylamino, thioether, sulfonylamino, nitro,halogen, CF₃, or cyano; and R₂′ is selected from lower alkyl, alkoxy,carboxyl, ester, amido, sulfonamido, heterocyclyl, cycloalkyl, hydroxyl,amino, acylamino, thioether, sulfonylamino, halogen, CF₃, or cyano.

In certain preferred embodiments, R₁ is independently selected from H,lower alkyl, or alkoxy, or R₁ taken together with the carbon to which itis attached forms a carbonyl.

In certain preferred embodiments, R₂ is absent or each R₂ isindependently selected from lower alkyl, alkoxy, or halogen.

In certain preferred embodiments, R₂′ is selected from lower alkyl,alkoxy, or halogen.

In certain preferred embodiments, R₃ is absent or each R₃ isindependently selected from lower alkyl, alkoxy, CF₃, nitro, or halogen.

In certain preferred embodiments, R₁, R₂, and R₃ are absent.

In certain embodiments, the TRPV3 antagonist can be represented byFormula VIII or a salt, solvate, hydrate, oxidative metabolite orprodrug thereof:

wherein: X represents O, S, N(CN), CF₂, or C(CN)₂; G represents N orC(R₃); R₁ and R₂, each independently, is absent or represents one ormore substituents on the ring to which it is attached; Ar′ represents anaryl or heteroaryl group; n represents 1, 2, or 3; R₃ represents,independently for each occurrence, H or a substituent; R₅ representsCH₂, O, NR₆; R₆ represents H or lower alkyl; and D represents O or N.

In certain embodiments, Ar′ represents phenyl, thiophene, imidazole,thiazole, furan, oxazole, tetrazole, thiadiazole, pyridine, pyrimadine,or triazole.

In certain embodiments, X represents S and D represents O.

In certain embodiments, R₁ is absent or, independently for eachoccurrence, is selected from lower alkyl, alkoxy, carboxyl, ester,amido, sulfonamido, heterocyclyl, cycloalkyl, hydroxyl, amino,acylamino, thioether, sulfonylamino, or R₁ taken together with thecarbon to which it is attached forms a carbonyl or thiocarbonyl; and R₂and R₃, independently for each occurrence, are selected from H, loweralkyl, alkoxy, carboxyl, ester, amido, sulfonamido, heterocyclyl,cycloalkyl, hydroxyl, amino, acylamino, thioether, sulfonylamino, nitro,halogen, CF₃, or cyano.

In certain embodiments, the TRPV3 antagonist can be represented byFormula VIIIa:

wherein: R₁, R₂ and R₃ are absent or represent one or more substituentson the ring to which they are attached.

In certain embodiments, R₁ is absent or, independently for eachoccurrence, is selected from lower alkyl, alkoxy, carboxyl, ester,amido, sulfonamido, heterocyclyl, cycloalkyl, hydroxyl, amino,acylamino, thioether, sulfonylamino, or R₁ taken together with thecarbon to which it is attached forms a carbonyl or thiocarbonyl; and R₂and R₃ are absent or, independently for each occurrence, are selectedfrom lower alkyl, alkoxy, carboxyl, ester, amido, sulfonamido,heterocyclyl, cycloalkyl, hydroxyl, amino, acylamino, thioether,sulfonylamino, nitro, halogen, CF₃, or cyano.

In certain embodiments, the TRPV3 antagonist is a compound of Formula IXor a salt, solvate, hydrate, oxidative metabolite or prodrug thereof:

wherein: A represents N or C(R₂); G represents N or C(R₃); R₁ representsH or a substituent; R₂ represents, independently for each occurrence, Hor a substituent; R₃ represents, independently for each occurrence, H ora substituent; R₃′ represents H, halogen, alkoxy, lower alkyl,perfluoroalkyl, aryl, heteroaryl, or CHR_(3a); R₃″ represents H,halogen, lower alkyl, aryl, heteroaryl, or CHR_(3a); R_(3a) representsamino, heterocyclyl, carboxyl, ester, aryl, or heteroaryl; and Drepresents O or N, provided that when R₃′ is H, R³″ is not H or loweralkyl, and when R₃′ is halogen, R₃″ is not H.

In certain embodiments, R₁ is selected from H, lower alkyl, alkoxy,carboxyl, ester, amido, sulfonamido, heterocyclyl, cycloalkyl, hydroxyl,amino, acylamino, thioether, sulfonylamino, or R₁ taken together withthe carbon to which it is attached forms a carbonyl or thiocarbonyl; andR₂ and R₃, independently for each occurrence, are selected from H, loweralkyl, alkoxy, carboxyl, ester, amido, sulfonamido, heterocyclyl,cycloalkyl, hydroxyl, amino, acylamino, thioether, sulfonylamino,halogen, CF₃, or cyano.

In certain embodiments, the TRPV3 antagonist is a compound can berepresented by formula IXa

wherein R₁ represents H or a substituent; R₂ is absent or represents asubstituent; R₃ is absent or represents a substituent; R₃′ represents H,halogen, alkoxy, lower alkyl, perfluoroalkyl, aryl, heteroaryl, orCHR_(3a); R₃″ represents H, halogen, lower alkyl, aryl, heteroaryl, orCHR_(3a); R_(3a) represents amino, heterocyclyl, carboxyl, ester, aryl,or heteroaryl; and D represents 0 or N, provided that when R₃′ is H, R₃″is not H or lower alkyl, and when R₃′ is halogen, R₃″ is not H.

In certain embodiments, R₁ is selected from H, lower alkyl, alkoxy,carboxyl, ester, amido, sulfonamido, heterocyclyl, cycloalkyl, hydroxyl,amino, acylamino, thioether, sulfonylamino, or R₁ taken together withthe carbon to which it is attached forms a carbonyl or thiocarbonyl; andR₂ and R₃ are absent or, independently for each occurrence, are selectedfrom lower alkyl, alkoxy, carboxyl, ester, amido, sulfonamido,heterocyclyl, cycloalkyl, hydroxyl, amino, acylamino, thioether,sulfonylamino, halogen, CF₃, or cyano.

In certain embodiments, the TRPV3 antagonist is a compound of Formula Xor a salt, solvate, hydrate, oxidative metabolite or prodrug thereof:

wherein: X represents O, S, N(CN), CF₂, or C(CN)₂; Z represents O, S,N(CN), CF₂, or C(CN)₂; A represents N or C(R₂); G represents N or C(R₃);R₁ represents H or a substituent; R₂ represents, independently for eachoccurrence, H or a substituent; R₃ represents, independently for eachoccurrence, H or a substituent; and D represents O or N.

In certain embodiments, X represents S and D represents O.

In certain embodiments, R₁ is selected from H, lower alkyl, alkoxy,carboxyl, ester, amido, sulfonamido, heterocyclyl, cycloalkyl, hydroxyl,amino, acylamino, thioether, sulfonylamino, or R₁ taken together withthe carbon to which it is attached forms a carbonyl or thiocarbonyl; andR₂ and R₃, independently for each occurrence, are selected from H, loweralkyl, alkoxy, carboxyl, ester, amido, sulfonamido, heterocyclyl,cycloalkyl, hydroxyl, amino, acylamino, thioether, sulfonylamino, nitro,halogen, CF₃, or cyano.

In certain embodiments, the TRPV3 antagonist is a compound can berepresented by Formula Xa:

wherein: R₁ represents H or a substituent; and R₂ and R₃ are absent orrepresent one or more substituents on the ring to which they areattached.

In certain embodiments, R₁ is selected from H, lower alkyl, alkoxy,carboxyl, ester, amido, sulfonamido, heterocyclyl, cycloalkyl, hydroxyl,amino, acylamino, thioether, sulfonylamino, or R₁ taken together withthe carbon to which it is attached forms a carbonyl or thiocarbonyl; andR₂ and R₃ are absent or, independently for each occurrence, are selectedfrom lower alkyl, alkoxy, carboxyl, ester, amido, sulfonamido,heterocyclyl, cycloalkyl, hydroxyl, amino, acylamino, thioether,sulfonylamino, nitro, halogen, CF₃, or cyano.

In certain embodiments, R₁ is H; and R₂ and R₃ are absent.

In certain embodiments, the TRPV3 antagonist is a compound representedby Formula Xb:

wherein: R₁ represents H or a substituent; and R₂ and R₃ are absent orrepresent one or more substituents on the ring to which they areattached.

In certain embodiments, wherein R₁ is selected from H, lower alkyl,alkoxy, carboxyl, ester, amido, sulfonamido, heterocyclyl, cycloalkyl,hydroxyl, amino, acylamino, thioether, sulfonylamino, or R₁ takentogether with the carbon to which it is attached forms a carbonyl orthiocarbonyl; and R₂ and R₃ are absent or, independently for eachoccurrence, are selected from lower alkyl, alkoxy, carboxyl, ester,amido, sulfonamido, heterocyclyl, cycloalkyl, hydroxyl, amino,acylamino, thioether, sulfonylamino, nitro, halogen, CF₃, or cyano.

In certain embodiments, the TRPV3 antagonist is a compound of Formula XIor a salt, solvate, hydrate, oxidative metabolite or prodrug thereof:

wherein: X represents O, S, N(CN), CF₂, or C(CN)₂; Z represents O, S,N(CN), CF₂, or C(CN)₂; A represents N or C(R₂); G represents N or C(R₃);R₁ represents H or a substituent; R₂ represents, independently for eachoccurrence, H or a substituent; R₃ represents, independently for eachoccurrence, H or a substituent; R_(3b) represents a substituent; and Drepresents O or N.

In certain embodiments, X represents S and D represents O.

In certain embodiments, R₁ is selected from H, lower alkyl, alkoxy,carboxyl, ester, amido, sulfonamido, heterocyclyl, cycloalkyl, hydroxyl,amino, acylamino, thioether, sulfonylamino, or R₁ taken together withthe carbon to which it is attached forms a carbonyl or thiocarbonyl; andR₂ and R₃ are absent or, independently for each occurrence, are selectedfrom lower alkyl, alkoxy, carboxyl, ester, amido, sulfonamido,heterocyclyl, cycloalkyl, hydroxyl, amino, acylamino, thioether,sulfonylamino, nitro, halogen, CF₃, or cyano; and R_(3b) is selectedfrom lower alkyl, alkoxy, carboxyl, ester, amido, sulfonamido,heterocyclyl, cycloalkyl, hydroxyl, amino, acylamino, thioether,sulfonylamino, halogen, CF₃, or cyano.

In certain embodiments, the TRPV3 antagonist is a compound representedby Formula XIa:

wherein: R₁ represents H or a substituent; R₂ and R₃ are absent orrepresent one or more substituents on the ring to which they areattached; and R_(3b) represents a substituent.

In certain embodiments, R₁ is selected from H, lower alkyl, alkoxy,carboxyl, ester, amido, sulfonamido, heterocyclyl, cycloalkyl, hydroxyl,amino, acylamino, thioether, sulfonylamino, or R₁ taken together withthe carbon to which it is attached forms a carbonyl or thiocarbonyl; R₂and R₃ are absent or, independently for each occurrence, are selectedfrom lower alkyl, alkoxy, carboxyl, ester, amido, sulfonamido,heterocyclyl, cycloalkyl, hydroxyl, amino, acylamino, thioether,sulfonylamino, nitro, halogen, CF₃, or cyano; and R_(3b) is selectedfrom lower alkyl, alkoxy, carboxyl, ester, amido, sulfonamido,heterocyclyl, cycloalkyl, hydroxyl, amino, acylamino, thioether,sulfonylamino, halogen, CF₃, or cyano.

In certain embodiments, R₁ is H; R₂ and R₃ are absent; and R_(3b) is ahalogen.

In certain embodiments, the TRPV3 antagonist is a compound can berepresented by Formula XIb:

wherein: R₁ represents H or a substituent; R₂ and R₃ are absent orrepresent one or more substituents on the ring to which they areattached; and R_(3b) represents a substituent.

In certain embodiments, R₁ is selected from H, lower alkyl, alkoxy,carboxyl, ester, amido, sulfonamido, heterocyclyl, cycloalkyl, hydroxyl,amino, acylamino, thioether, sulfonylamino, or R₁ taken together withthe carbon to which it is attached forms a carbonyl or thiocarbonyl; R₂and R₃ are absent or, independently for each occurrence, are selectedfrom lower alkyl, alkoxy, carboxyl, ester, amido, sulfonamido,heterocyclyl, cycloalkyl, hydroxyl, amino, acylamino, thioether,sulfonylamino, nitro, halogen, CF₃, or cyano; and R_(3b) is selectedfrom lower alkyl, alkoxy, carboxyl, ester, amido, sulfonamido,heterocyclyl, cycloalkyl, hydroxyl, amino, acylamino, thioether,sulfonylamino, halogen, CF₃, or cyano.

In certain embodiments of the above formulae, substituents may includeone or more of: alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl,heteroaryl, cycloalkylalkyl, heterocyclylalkyl, aralkyl, orheteroaralkyl, any of which may itself be further substituted, orhalogen, carbonyl (e.g., ester, carboxyl, or formyl), thiocarbonyl(e.g., thioester, thiocarboxylate, or thioformate), ketone, aldehyde, ahydroxyl, an alkoxyl, a sulfhydryl, an alkylthio, amino, acylamino,amido, amidino, cyano, nitro, azido, sulfonyl, sulfoxido, sulfate,sulfonate, sulfamoyl, sulfonamido, a phosphate, and a phosphoryl.

Compounds of any of the above structures may be used in the manufactureof medicaments for the treatment of any diseases disclosed herein.Furthermore, TRPV3 inhibitors, according to the present invention, canbe used in the manufacture of medicaments for the treatment of anydiseases disclosed herein. In certain embodiments, compounds (e.g.,inhibitors) of the invention can be used in the treatment of pain.

Compounds of any of the above structures may be used to inhibit anactivity of TRPV3 in vitro or in vivo, and/or can be used in themanufacture of medicaments to inhibit an activity of TRPV3 in vitro orin vivo. TRPV3 inhibitors, according to the present invention, can beused to inhibit an activity of TRPV3, and/or can be used in themanufacture of medicaments to inhibit an activity of TRPV3 in vitro orin vivo.

For any of the aspects or embodiments of the invention, an exemplaryfunction of TRPV3 that may be inhibited or modulated by a compound ofthe invention is a TRPV3-mediated current (e.g., an inward or outwardPhase I and/or Phase II current).

In particular embodiments, the small molecule is chosen for use becauseit is more selective for one TRP isoform than others, e.g., 10-fold, andmore preferably at least 100- or even 1000-fold more selective for TRPV3over one or more of TRPC6, TRPV5, TRPV6, TRPM8, TRPV1, and/or TRPV4. Inother embodiments, the differential is smaller, e.g., it more stronglyinhibits TRPV3 than TRPM8, TRPV1 and/or TRPV4, preferably at leasttwice, three times, five times, or even ten times more strongly. Suchcomparisons may be made, for example, by comparing IC₅₀ values.

In certain embodiments, a compound which is an antagonist of TRPV3 ischosen to selectively antagonize TRPV3 over other ion channels, e.g.,the compound modulates the activity of TRPV3 at least an order ofmagnitude more strongly than it modulates the activity of NaV1.2,Cav1.2, Cav3.1, HERG, and/or mitochondrial uniporter, preferably atleast two orders of magnitude more strongly, even more preferably atleast three orders of magnitude more strongly. Such comparisons may bemade, for example, by comparing IC₅₀ values.

Similarly, in particular embodiments, the small molecule is chosen foruse because it lacks significant activity against one or more targetsother than TRPV3. For example, the compound may have an IC₅₀ above 500nM, above 1 μM, or even above 10 μM for inhibiting one or more of TRPC6,TRPV5, TRPV6, TRPV1, NaV1.2, Cav1.2, Cav3.1, HERG, and the mitochondrialuniporter.

In certain embodiment, the small molecule is chosen because itantagonizes the function of both TRPV3 and TRPM8, TRPV1 and/or TRPV4.Although such compounds selectively antagonize the function of both ionchannels, the IC₅₀ values need not be identical.

In certain embodiments of any of the foregoing, the small molecule maybe chosen because it is capable of inhibiting heat-induced activation ofTRPV3. In certain embodiments, the TRPV3 antagonist inhibitsheat-induced activation of TRPV3 and 2-APB induced activation of TRPV3.In certain other embodiments, the TRPV3 antagonist inhibits heat-inducedactivation of TRPV3 but does not inhibit 2-APB induced activation ofTRPV3.

In certain embodiments of any of the foregoing, the small molecule maybe chosen because it inhibits a TRPV3 function with an IC₅₀ less than 1uM, or even less than 700, 600, 500, 400, 300, 200, or 100 nM. In otherembodiments, the small molecule is chosen because it inhibits a TRPV3function with an IC₅₀ less than 50 nM, or even less than 25, 10, or 1nM.

In certain embodiments of any of the foregoing, the compound may bechosen based on the rate of inhibition of a TRPV3 function. In oneembodiment, the compound inhibits a TRPV3 function in less than 5minutes, preferably less than 4, 3, or 2 minutes. In another embodiment,the compound inhibits a TRPV3 function in less than 1 minute.

In any of the foregoing embodiments, the small molecule antagonist ofTRPV3 function may inhibit the Phase I outward current, the Phase Iinward current, the Phase II outward current, the Phase II inwardcurrent, or any combination of one or more of these currents. Compoundsthat inhibit more than one of the foregoing currents may do so with thesame or with differing IC₅₀ values. In any of the foregoing, the abilityof a compound to inhibit a particular Phase I and/or Phase II currentcan be assessed either in vitro or in vivo. Compounds that inhibit anyof the foregoing currents in an in vitro or in vivo assay arecharacterized as compounds that inhibit a function of TRPV3.

In certain embodiments of any of the foregoing, inhibition of a TRPV3function means that a function, for example a TRPV3 mediated current, isdecreased by at least 25%, 30%, 40%, or 50% in the presence of aneffective amount of a compound in comparison to in the absence of thecompound or in comparison to an ineffective amount of a compound. Incertain other embodiments, the inhibition of a TRPV3 function means thata function, for example a TRPV3 mediated current, is decreased by atleast 50%, 60%, 70%, 75%, 80%, 85%, or 90% in the presence of aneffective amount of a compound in comparison to in the absence of thecompound. In still other embodiments, the inhibition of a TRPV3 functionmeans that a function, for example a TRPV3 mediated current, isdecreased by at least 92%, 95%, 97%, 98%, 98%, 99%, or 100% in thepresence of an effective amount of a compound in comparison to in theabsence of the compound.

In certain embodiments of any of the foreging, the TRPV3 inhibitor isused to treat or ameliorate pain. Exemplary classes of pain that cantreated using a TRPV3 inhibitor include, but are not limited tonociceptive pain, inflammatory pain, and neuropathic pain. Pain that canbe treated with a TRPV3 inhibitor can be chronic or acute.

In certain embodiments of any of the foregoing, the TRPV3 inhibitor canbe used to treat or ameliorate pain with fewer side effects. Forexample, the TRPV3 inhibitor can be used to treat or ameliorate painwithout the narcotic effects of, for example, morphine.

In any of the foregoing embodiments, IC₅₀ values are measured in vitrousing, for example, patch clamp analysis or standard measurements ofcalcium flux. Exemplary in vitro methods that can be used to measureIC₅₀ values of a compound are described in Examples 2 and 3.

Without being bound by theory, a compound may inhibit a function ofTRPV3 by binding covalently or non-covalently to a portion of TRPV3.Alternatively, a compound may inhibit a function of TRPV3 indirectly,for example, by associating with a protein or non-protein cofactornecessary for a function of TRPV3. One of skill in the art will readilyappreciate that an inhibitory compound may associate reversibly orirreversibly with TRPV3 or a cofactor thereof. Compounds that reversiblyassociate with TRPV3 or a cofactor thereof may continue to inhibit afunction of TRPV3 even after dissociation.

The subject TRPV3 inhibitors can be used alone or in combination withother pharmaceutically active agents. Examples of such otherpharmaceutically active agents include, but are not limited to,anti-inflammatory agents (e.g., NSAIDS, hormones and autacoids such ascorticosteroids), anti-acne agents (e.g., retinoids), anti-wrinkleagents, anti-scarring agents, anti-psoriatic agents, anti-proliferativeagents (e.g., anti-eczema agents), anti-fungal agents, anti-viralagents, anti-septic agents (e.g., antibacterials), local anaesthetics,anti-migraine agents, keratolytic agents, hair growth stimulants, hairgrowth inhibitors, and other agents used for the treatment of skindiseases or conditions. Certain active agents belong to more than onecategory.

The subject TRPV3 inhibitors can be used alone or as part of atherapeutic regimen combined with other treatments, therapies, orinterventions appropriate for the particular disease, condition, injuryor disorder being treated. When used as part of a therapeutic regimen,the invention contemplates use of TRPV3 inhibitors in combination withone or more of the following treatment modalities: administration ofnon-TRPV3 inhibitor pharmaceuticals, chemotherapy, radiotherapy,homeopathic therapy, diet, stress management, and surgery.

When administered alone or as part of a therapeutic regimen, in certainembodiments, the invention contemplates administration of TRPV3inhibitors to treat a particular primary disease, injury, disorder, orcondition. Additionally or alternatively, the invention contemplatesadministration of TRPV3 inhibitors to treat pain associated with adisease, injury, disorder, or condition. In still other embodiments, theinvention contemplates administration of TRPV3 inhibitors to treatsymptoms secondary to the primary disease, injury, disorder, orconditions.

Definitions

The terms “antagonist” and “inhibitor” are used interchangeably to referto an agent that decreases or suppresses a biological activity, such asto repress an activity of an ion channel, such as TRPV3.

An “effective amount” of, e.g., a TRPV3 antagonist, with respect to thesubject method of treatment, refers to an amount of the antagonist in apreparation which, when applied as part of a desired dosage regimenbrings about a desired clinical or functional result. Without beingbound by theory, an effective amount of a TRPV3 antagonist for use inthe methods of the present invention, includes an amount of a TRPV3antagonist effective to decrease one or more in vitro or in vivofunction of a TRPV3 channel. Exemplary functions include, but are notlimited to, intracellular calcium levels, membrane polarization (e.g.,an antagonist may promote hyperpolarization of a cell), Phase I outwardcurrent, Phase II outward current, Phase I inward current, and Phase IIinward current. Compounds that antagonize TRPV3 function includecompounds that antagonize an in vitro or in vivo functional activity ofTRPV3. When a particular functional activity is only readily observablein an in vitro assay, the ability of a compound to inhibit TRPV3function in that in vitro assay serves as a reasonable proxy for theactivity of that compound.

The term “preventing” is art-recognized, and when used in relation to acondition, such as a local recurrence (e.g., pain), a disease such ascancer, a syndrome complex such as heart failure or any other medicalcondition, is well understood in the art, and includes administration ofa composition which reduces the frequency of, or delays the onset of,symptoms of a medical condition in a subject relative to a subject whichdoes not receive the composition. Thus, prevention of cancer includes,for example, reducing the number of detectable cancerous growths in apopulation of patients receiving a prophylactic treatment relative to anuntreated control population, and/or delaying the appearance ofdetectable cancerous growths in a treated population versus an untreatedcontrol population, e.g., by a statistically and/or clinicallysignificant amount. Prevention of an infection includes, for example,reducing the number of diagnoses of the infection in a treatedpopulation versus an untreated control population, and/or delaying theonset of symptoms of the infection in a treated population versus anuntreated control population. Prevention of pain includes, for example,reducing the magnitude of, or alternatively delaying, pain sensationsexperienced by subjects in a treated population versus an untreatedcontrol population.

The present invention provides compounds which are in prodrug form. Theterm “prodrug” is intended to encompass compounds that, underphysiological conditions, are converted into the therapeutically activeagents of the present invention. A common method for making a prodrug isto include selected moieties that are hydrolyzed under physiologicalconditions to reveal the desired molecule. In other embodiments, theprodrug is converted by an enzymatic activity of the host animal.Additionally, prodrugs can be converted to the compounds of the presentinvention by chemical or biochemical methods in an ex vivo environment.For example, prodrugs can be slowly converted to the compounds of thepresent invention when placed in a transdermal patch reservoir with asuitable enzyme or chemical reagent.

The term “treating” includes prophylactic and/or therapeutic treatments.The term “prophylactic or therapeutic” treatment is art-recognized andincludes administration to the host of one or more of the subjectcompositions. If it is administered prior to clinical manifestation ofthe unwanted condition (e.g., disease or other unwanted state of thehost animal) then the treatment is prophylactic, (i.e., it protects thehost against developing the unwanted condition), whereas if it isadministered after manifestation of the unwanted condition, thetreatment is therapeutic, (i.e., it is intended to diminish, ameliorate,or stabilize the existing unwanted condition or side effects thereof).

The terms “TRPV3”, “TRPV3 protein”, and “TRPV3 channel” are usedinterchangeably throughout the application. These terms refer to an ionchannel (e.g., a polypeptide) comprising the amino acid sequence, forexample, the amino acid sequence of a human TRPV3 protein, or anequivalent polypeptide, or a functional bioactive fragment thereof. Incertain embodiments, the term refers to a polypeptide comprising,consisting of, or consisting essentially of, a TRPV3 amino acid sequenceset forth, for example, in any of the patent applications referencedherein. TRPV3 protein may also include orthologs, for example, mouse,rat, horse, or Drosophila TRPV3.

TRPV3 includes polypeptides that retain a function of TRPV3 and comprise(i) all or a portion of a TRPV3 amino acid sequence (ii) a TRPV3 aminoacid sequence with 1 to about 2, 3, 5, 7, 10, 15, 20, 30, 50, 75 or moreconservative amino acid substitutions; (iii) an amino acid sequence thatis at least 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99%identical to a TRPV3 amino acid sequence; and (iv) functional fragmentsthereof. Polypeptides of the invention also include homologs, e.g.,orthologs and paralogs, of a human TRPV3 polypeptide. TRPV3 polypeptidesand amino acid sequences include, for example, the sequences set forthin any of the patent applications referenced herein.

The term “TRPV3” further refers to a nucleic acid encoding a polypeptideof the invention, e.g., a nucleic acid comprising a sequence consistingof, or consisting essentially of, a TRPV3 polynucleotide sequence. Anucleic acid of the invention may comprise all, or a portion of: (i) aTRPV3 nucleotide sequence; (ii) a nucleotide sequence at least 70%, 75%,80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% identical to a TRPV3 nucleotidesequence; (iii) a nucleotide sequence that hybridizes under stringentconditions to a TRPV3 nucleotide sequence; (iv) nucleotide sequencesencoding polypeptides that are functionally equivalent to polypeptidesof the invention; (v) nucleotide sequences encoding polypeptides atleast about 70%, 75%, 80%, 85%, 90%, 95%, 98%, 99% homologous oridentical with a TRPV3 polypeptide sequence; (vi) nucleotide sequencesencoding polypeptides having an activity of a polypeptide of theinvention and having at least about 70%, 75%, 80%, 85%, 90%, 95%, 98%,99% or more homology or identity with a TRPV3 polypeptide sequence;(vii) nucleotide sequences that differ by 1 to about 2, 3, 5, 7, 10, 15,20, 30, 50, 75 or more nucleotide substitutions, additions or deletions,such as allelic variants, of a TRPV3 nucleotide sequence; (viii) nucleicacids derived from and evolutionarily related to a TRPV3 nucleotidesequence; and (ix) complements of, and nucleotide sequences resultingfrom the degeneracy of the genetic code, for all of the foregoing andother nucleic acids of the invention. Nucleic acids of the inventionalso include homologs, e.g., orthologs and paralogs, of a TRPV3 nucleicacid sequence and also variants which have been codon optimized forexpression in a particular organism (e.g., host cell). TRPV3 nucleicacid sequences include, for example, the sequences set forth in any ofthe patent applications referenced herein. Where not explicitly stated,one of skill in the art can readily assess whether TRPV3 refers to anucleic acid or a protein.

The terms “compound” and “agent” are used interchangeably to refer tothe inhibitors/antagonists of the invention. In certain embodiments, thecompounds are small organic or inorganic molecules, e.g., with molecularweights less than 7500 amu, preferably less than 5000 amu, and even morepreferably less than 2000, 1500, 1000, or 500 amu. One class of smallorganic or inorganic molecules are non-peptidyl, e.g., containing 2, 1,or no peptide and/or saccharide linkages. In certain other embodiments,the compounds are proteins, for example, antibodies or aptamers. Suchcompounds can bind to and inhibit a function of TRPV3. In certain otherembodiments, the compounds are nucleic acids, for example, TRPV3antisense oligonucleotides or TRPV3 RNAi constructs. Such compounds caninhibit the expression of TRPV3, thereby inhibiting the activity ofTRPV3. Other exemplary compounds that may act as inhibitors includeribozymes and peptide fragments.

The term “acylamino” is art-recognized and refers to a moiety that canbe represented by the general formula:

wherein R₉ is as defined above, and R′₁₁ represents a hydrogen, analkyl, an alkenyl or —(CH₂)_(m)—R₈, where m and R₈ are as defined above.

Herein, the term “aliphatic group” refers to a straight-chain,branched-chain, or cyclic aliphatic hydrocarbon group and includessaturated and unsaturated aliphatic groups, such as an alkyl group, analkenyl group, and an alkynyl group.

The terms “alkenyl” and “alkynyl” refer to unsaturated aliphatic groupsanalogous in length and possible substitution to the alkyls describedabove, but that contain at least one double or triple bond respectively.

The terms “alkoxyl” or “alkoxy” as used herein refers to an alkyl group,as defined below, having an oxygen radical attached thereto.Representative alkoxyl groups include methoxy, ethoxy, propyloxy,tert-butoxy and the like. An “ether” is two hydrocarbons covalentlylinked by an oxygen. Accordingly, the substituent of an alkyl thatrenders that alkyl an ether is or resembles an alkoxyl, such as can berepresented by one of —O-alkyl, —O-alkenyl, —O-alkynyl, —O—(CH₂)_(m)—R₈,where m and R₈ are described above.

The term “alkyl” refers to the radical of saturated aliphatic groups,including straight-chain alkyl groups, branched-chain alkyl groups,cycloalkyl (alicyclic) groups, alkyl-substituted cycloalkyl groups, andcycloalkyl-substituted alkyl groups. In preferred embodiments, astraight chain or branched chain alkyl has 30 or fewer carbon atoms inits backbone (e.g., C₁-C₃₀ for straight chains, C₃-C₃₀ for branchedchains), and more preferably 20 or fewer, and most preferably 10 orfewer. Likewise, preferred cycloalkyls have from 3-10 carbon atoms intheir ring structure, and more preferably have 5, 6 or 7 carbons in thering structure.

Moreover, the term “alkyl” (or “lower alkyl”) as used throughout thespecification, examples, and claims is intended to include both“unsubstituted alkyls” and “substituted alkyls”, the latter of whichrefers to alkyl moieties having substituents replacing a hydrogen on oneor more carbons of the hydrocarbon backbone. Such substituents caninclude, for example, a halogen, a hydroxyl, a carbonyl (such as acarboxyl, an alkoxycarbonyl, a formyl, or an acyl), a thiocarbonyl (suchas a thioester, a thioacetate, or a thioformate), an alkoxyl, aphosphoryl, a phosphate, a phosphonate, a phosphinate, an amino, anamido, an amidine, an imine, a cyano, a nitro, an azido, a sulfhydryl,an alkylthio, a sulfate, a sulfonate, a sulfamoyl, a sulfonamido, asulfonyl, a heterocyclyl, an aralkyl, or an aromatic or heteroaromaticmoiety. It will be understood by those skilled in the art that themoieties substituted on the hydrocarbon chain can themselves besubstituted, if appropriate. For instance, the substituents of asubstituted alkyl may include substituted and unsubstituted forms ofamino, azido, imino, amido, phosphoryl (including phosphonate andphosphinate), sulfonyl (including sulfate, sulfonamido, sulfamoyl andsulfonate), and silyl groups, as well as ethers, alkylthios, carbonyls(including ketones, aldehydes, carboxylates, and esters), —CF₃, —CN andthe like. Exemplary substituted alkyls are described below. Cycloalkylscan be further substituted with alkyls, alkenyls, alkoxys, alkylthios,aminoalkyls, carbonyl-substituted alkyls, —CF₃, —CN, and the like.

Analogous substitutions can be made to alkenyl and alkynyl groups toproduce, for example, aminoalkenyls, aminoalkynyls, amidoalkenyls,amidoalkynyls, iminoalkenyls, iminoalkynyls, thioalkenyls, thioalkynyls,carbonyl-substituted alkenyls or alkynyls.

Unless the number of carbons is otherwise specified, “lower alkyl” asused herein means an alkyl group, as defined above, but having from oneto ten carbons, more preferably from one to six carbon atoms in itsbackbone structure. Likewise, “lower alkenyl” and “lower alkynyl” havesimilar chain lengths. Throughout the application, preferred alkylgroups are lower alkyls. In preferred embodiments, a substituentdesignated herein as alkyl is a lower alkyl.

The term “alkylthio” refers to an alkyl group, as defined above, havinga sulfur radical attached thereto. In preferred embodiments, the“alkylthio” moiety is represented by one of —S-alkyl, —S-alkenyl,—S-alkynyl, and —S—(CH₂)_(m)—R₈, wherein m and R₈ are defined above.Representative alkylthio groups include methylthio, ethylthio, and thelike.

The terms “amine” and “amino” are art-recognized and refer to bothunsubstituted and substituted amines, e.g., a moiety that can berepresented by the general formula:

wherein R₉, R₁₀ and R′₁₀ each independently represent a hydrogen, analkyl, an alkenyl, —(CH₂)_(m)—R₈, or R₉ and R₁₀ taken together with theN atom to which they are attached complete a heterocycle having from 4to 8 atoms in the ring structure; R₈ represents an aryl, a cycloalkyl, acycloalkenyl, a heterocycle or a polycycle; and m is zero or an integerin the range of 1 to 8. In preferred embodiments, only one of R₉ or R₁₀can be a carbonyl, e.g., R₉, R₁₀ and the nitrogen together do not forman imide. In certain such embodiments, neither R₉ and R₁₀ is attached toN by a carbonyl, e.g., the amine is not an amide or imide, and the amineis preferably basic, e.g., its conjugate acid has a pK_(a) above 7. Ineven more preferred embodiments, R₉ and R₁₀ (and optionally R′₁₀) eachindependently represent a hydrogen, an alkyl, an alkenyl, or—(CH₂)_(m)—R₈. Thus, the term “alkylamine” as used herein means an aminegroup, as defined above, having a substituted or unsubstituted alkylattached thereto, i.e., at least one of R₉ and R₁₀ is an alkyl group.

The term “amido” is art-recognized as an amino-substituted carbonyl andincludes a moiety that can be represented by the general formula:

wherein R₉, R₁₀ are as defined above. Preferred embodiments of the amidewill not include imides that may be unstable.

The term “aralkyl”, as used herein, refers to an alkyl group substitutedwith an aryl group (e.g., an aromatic or heteroaromatic group).

The term “aryl” as used herein includes 5-, 6-, and 7-memberedsingle-ring aromatic groups that may include from zero to fourheteroatoms, for example, benzene, pyrrole, furan, thiophene, imidazole,oxazole, thiazole, triazole, pyrazole, pyridine, pyrazine, pyridazineand pyrimidine, and the like. Those aryl groups having heteroatoms inthe ring structure may also be referred to as “aryl heterocycles” or“heteroaromatics.” The aromatic ring can be substituted at one or morering positions with such substituents as described above, for example,halogen, azide, alkyl, aralkyl, alkenyl, alkynyl, cycloalkyl, hydroxyl,alkoxyl, amino, nitro, sulfhydryl, imino, amido, phosphate, phosphonate,phosphinate, carbonyl, carboxyl, silyl, ether, alkylthio, sulfonyl,sulfonamido, ketone, aldehyde, ester, heterocyclyl, aromatic orheteroaromatic moieties, —CF₃, —CN, or the like. The term “aryl” alsoincludes polycyclic ring systems having two or more cyclic rings inwhich two or more carbons are common to two adjoining rings (the ringsare “fused rings”) wherein at least one of the rings is aromatic, e.g.,the other cyclic rings can be cycloalkyls, cycloalkenyls, cycloalkynyls,aryls and/or heterocyclyls.

The term “carbocycle”, as used herein, refers to an aromatic ornon-aromatic ring in which each atom of the ring is carbon.

The term “carbonyl” is art-recognized and includes such moieties as canbe represented by the general formula:

wherein X is a bond or represents an oxygen or a sulfur, and R₁₁represents a hydrogen, an alkyl, an alkenyl, —(CH₂)_(m)—R₈ or apharmaceutically acceptable salt, R′₁₁ represents a hydrogen, an alkyl,an alkenyl or —(CH₂)_(m)—R₈, where m and R₈ are as defined above. WhereX is an oxygen and R₁₁ or R′₁₁ is not hydrogen, the formula representsan “ester”. Where X is an oxygen, and R₁₁ is as defined above, themoiety is referred to herein as a carboxyl group, and particularly whenR₁₁ is a hydrogen, the formula represents a “carboxylic acid”. Where Xis an oxygen, and R′₁₁ is hydrogen, the formula represents a “formate”.In general, where the oxygen atom of the above formula is replaced bysulfur, the formula represents a “thiocarbonyl” group. Where X is asulfur and R₁₁ or R′₁₁ is not hydrogen, the formula represents a“thioester.” Where X is a sulfur and R₁₁ is hydrogen, the formularepresents a “thiocarboxylic acid.” Where X is a sulfur and R₁₁′ ishydrogen, the formula represents a “thiolformate.” On the other hand,where X is a bond, and R₁₁ is not hydrogen, the above formula representsa “ketone” group. Where X is a bond, and R₁₁ is hydrogen, the aboveformula represents an “aldehyde” group.

The term “heteroatom” as used herein means an atom of any element otherthan carbon or hydrogen. Preferred heteroatoms are boron, nitrogen,oxygen, phosphorus, sulfur and selenium.

The terms “heterocyclyl” or “heterocyclic group” refer to 3- to10-membered ring structures, more preferably 3- to 7-membered rings,whose ring structures include one to four heteroatoms. Heterocycles canalso be polycycles. Heterocyclyl groups include, for example, thiophene,thianthrene, furan, pyran, isobenzofuran, chromene, xanthene,phenoxathiin, pyrrole, imidazole, pyrazole, isothiazole, isoxazole,pyridine, pyrazine, pyrimidine, pyridazine, indolizine, isoindole,indole, indazole, purine, quinolizine, isoquinoline, quinoline,phthalazine, naphthyridine, quinoxaline, quinazoline, cinnoline,pteridine, carbazole, carboline, phenanthridine, acridine, pyrimidine,phenanthroline, phenazine, phenarsazine, phenothiazine, furazan,phenoxazine, pyrrolidine, oxolane, thiolane, oxazole, piperidine,piperazine, morpholine, lactones, lactams such as azetidinones andpyrrolidinones, sultams, sultones, and the like. The heterocyclic ringcan be substituted at one or more positions with such substituents asdescribed above, as for example, halogen, alkyl, aralkyl, alkenyl,alkynyl, cycloalkyl, hydroxyl, amino, nitro, sulfhydryl, imino, amido,phosphate, phosphonate, phosphinate, carbonyl, carboxyl, silyl, ether,alkylthio, sulfonyl, ketone, aldehyde, ester, a heterocyclyl, anaromatic or heteroaromatic moiety, —CF₃, —CN, or the like.

As used herein, the term “nitro” means —NO₂; the term “halogen”designates —F, —Cl, —Br or —I; the term “sulfhydryl” means —SH; the term“hydroxyl” means —OH; and the term “sulfonyl” means —SO₂—.

The terms “polycyclyl” or “polycyclic group” refer to two or more rings(e.g., cycloalkyls, cycloalkenyls, cycloalkynyls, aryls and/orheterocyclyls) in which two or more carbons are common to two adjoiningrings, e.g., the rings are “fused rings”. Rings that are joined throughnon-adjacent atoms are termed “bridged” rings. Each of the rings of thepolycycle can be substituted with such substituents as described above,as for example, halogen, alkyl, aralkyl, alkenyl, alkynyl, cycloalkyl,hydroxyl, amino, nitro, sulfhydryl, imino, amido, phosphate,phosphonate, phosphinate, carbonyl, carboxyl, silyl, ether, alkylthio,sulfonyl, ketone, aldehyde, ester, a heterocyclyl, an aromatic orheteroaromatic moiety, —CF₃, —CN, or the like.

The phrase “protecting group” as used herein means temporarysubstituents which protect a potentially reactive functional group fromundesired chemical transformations. Examples of such protecting groupsinclude esters of carboxylic acids, silyl ethers of alcohols, andacetals and ketals of aldehydes and ketones, respectively. The field ofprotecting group chemistry has been reviewed (Greene, T. W.; Wuts, P. G.M. Protective Groups in Organic Synthesis, 2^(nd) ed.; Wiley: New York,1991).

As used herein, the term “substituted” is contemplated to include allpermissible substituents of organic compounds. In a broad aspect, thepermissible substituents include acyclic and cyclic, branched andunbranched, carbocyclic and heterocyclic, aromatic and nonaromaticsubstituents of organic compounds (e.g., alkyl, alkenyl, alkynyl,cycloalkyl, heterocyclyl, aryl, heteroaryl, cycloalkylalkyl,heterocyclylalkyl, aralkyl, or heteroaralkyl, any of which may itself befurther substituted), as well as halogen, carbonyl (e.g., ester,carboxyl, or formyl), thiocarbonyl (e.g., thioester, thiocarboxylate, orthioformate), ketone, aldehyde, amino, acylamino, amido, amidino, cyano,nitro, azido, sulfonyl, sulfoxido, sulfate, sulfonate, sulfamoyl,sulfonamido, and phosphoryl. Illustrative substituents include, forexample, those described herein above. The permissible substituents canbe one or more and the same or different for appropriate organiccompounds. For purposes of this invention, the heteroatoms such asnitrogen may have hydrogen substituents and/or any permissiblesubstituents of organic compounds described herein which satisfy thevalences of the heteroatoms. This invention is not intended to belimited in any manner by the permissible substituents of organiccompounds.

It will be understood that “substitution” or “substituted with” includesthe implicit proviso that such substitution is in accordance withpermitted valence of the substituted atom and the substituent, and thatthe substitution results in a stable compound, e.g., which does notspontaneously undergo transformation such as by rearrangement,cyclization, elimination, etc.

The term “sulfamoyl” is art-recognized and includes a moiety that can berepresented by the general formula:

in which R₉ and R₁₀ are as defined above.

The term “sulfate” is art recognized and includes a moiety that can berepresented by the general formula:

in which R₄₁ is as defined above.

The term “sulfonamido” is art recognized and includes a moiety that canbe represented by the general formula:

in which R₉ and R′₁₁ are as defined above.

The term “sulfonate” is art-recognized and includes a moiety that can berepresented by the general formula:

in which R₄₁ is an electron pair, hydrogen, alkyl, cycloalkyl, or aryl.

The terms “sulfoxido” or “sulfinyl”, as used herein, refers to a moietythat can be represented by the general formula:

in which R₄₄ is selected from the group consisting of hydrogen, alkyl,alkenyl, alkynyl, cycloalkyl, heterocyclyl, aralkyl, or aryl.

As used herein, the definition of each expression, e.g., alkyl, m, n,etc., when it occurs more than once in any structure, is intended to beindependent of its definition elsewhere in the same structure.

The terms triflyl, tosyl, mesyl, and nonaflyl are art-recognized andrefer to trifluoromethanesulfonyl, p-toluenesulfonyl, methanesulfonyl,and nonafluorobutanesulfonyl groups, respectively. The terms triflate,tosylate, mesylate, and nonaflate are art-recognized and refer totrifluoromethanesulfonate ester, p-toluenesulfonate ester,methanesulfonate ester, and nonafluorobutanesulfonate ester functionalgroups and molecules that contain said groups, respectively.

The abbreviations Me, Et, Ph, Tf, Nf, Ts, Ms represent methyl, ethyl,phenyl, trifluoromethanesulfonyl, nonafluorobutanesulfonyl,p-toluenesulfonyl and methanesulfonyl, respectively. A morecomprehensive list of the abbreviations utilized by organic chemists ofordinary skill in the art appears in the first issue of each volume ofthe Journal of Organic Chemistry; this list is typically presented in atable entitled Standard List of Abbreviations. The abbreviationscontained in said list, and all abbreviations utilized by organicchemists of ordinary skill in the art are hereby incorporated byreference.

Certain compounds of the present invention may exist in particulargeometric or stereoisomeric forms. The present invention contemplatesall such compounds, including cis- and trans-isomers, R- andS-enantiomers, diastereomers, (D)-isomers, (L)-isomers, the racemicmixtures thereof, and other mixtures thereof, as falling within thescope of the invention. Additional asymmetric carbon atoms may bepresent in a substituent such as an alkyl group. All such isomers, aswell as mixtures thereof, are intended to be included in this invention.

Methods of preparing substantially isomerically pure compounds are knownin the art. If, for instance, a particular enantiomer of a compound ofthe present invention is desired, it may be prepared by asymmetricsynthesis, or by derivation with a chiral auxiliary, where the resultingdiastereomeric mixture is separated and the auxiliary group cleaved toprovide the pure desired enantiomers. Alternatively, where the moleculecontains a basic functional group, such as amino, or an acidicfunctional group, such as carboxyl, diastereomeric salts may be formedwith an appropriate optically active acid or base, followed byresolution of the diastereomers thus formed by fractionalcrystallization or chromatographic means well known in the art, andsubsequent recovery of the pure enantiomers. Alternatively,enantiomerically enriched mixtures and pure enantiomeric compounds canbe prepared by using synthetic intermediates that are enantiomericallypure in combination with reactions that either leave the stereochemistryat a chiral center unchanged or result in its complete inversion.Techniques for inverting or leaving unchanged a particular stereocenter,and those for resolving mixtures of stereoisomers are well known in theart, and it is well within the ability of one of skill in the art tochoose an appropriate method for a particular situation. See, generally,Furniss et al. (eds.), Vogel's Encyclopedia of Practical OrganicChemistry 5^(th) Ed., Longman Scientific and Technical Ltd., Essex,1991, pp. 809-816; and Heller, Acc. Chem. Res. 23: 128 (1990).

Contemplated equivalents of the compounds described above includecompounds which otherwise correspond thereto, and which have the samegeneral properties thereof (e.g., the ability to inhibit TRPV3activity), wherein one or more simple variations of substituents aremade which do not adversely affect the efficacy of the compound. Ingeneral, the compounds of the present invention may be prepared by themethods illustrated in the general reaction schemes as, for example,described below, or by modifications thereof, using readily availablestarting materials, reagents and conventional synthesis procedures. Inthese reactions, it is also possible to make use of variants which arein themselves known, but are not mentioned here.

For purposes of this invention, the chemical elements are identified inaccordance with the Periodic Table of the Elements, CAS version,Handbook of Chemistry and Physics, 67th Ed., 1986-87, inside cover. Alsofor purposes of this invention, the term “hydrocarbon” is contemplatedto include all permissible compounds having at least one hydrogen andone carbon atom. In a broad aspect, the permissible hydrocarbons includeacyclic and cyclic, branched and unbranched, carbocyclic andheterocyclic, aromatic and nonaromatic organic compounds which can besubstituted or unsubstituted.

The compounds of the present invention may also contain unnaturalproportions of atomic isotopes at one or more of the atoms thatconstitute such compounds. For example, the compounds may beradiolabeled with radioactive isotopes, such as for example tritium(3H), iodine-125 (¹²⁵I) or carbon-14 (¹⁴C). All isotopic variations ofthe compounds of the present invention, whether radioactive or not, areintended to be encompassed within the scope of the present invention.

The symbol

, whether utilized as a bond or displayed perpendicular to a bondindicates the point at which the displayed moiety is attached to theremainder of the molecule, solid support, etc.

Certain compounds of the present invention can exist in unsolvated formsas well as solvated forms, including hydrated forms. In general, thesolvated forms are equivalent to unsolvated forms and are encompassedwithin the scope of the present invention. Certain compounds of thepresent invention may exist in multiple crystalline or amorphous forms.In general, all physical forms are equivalent for the uses contemplatedby the present invention and are intended to be within the scope of thepresent invention.

Wherein substituent groups are specified by their conventional chemicalformulae, written from left to right, they equally encompass thechemically identical substituents, which would result from writing thestructure from right to left, e.g., —CH₂O— is intended to also recite—OCH₂—; —NHS(O)₂— is also intended to represent —S(O)₂HN—; etc.

The term “pharmaceutically acceptable salts” includes salts of theactive compounds which are prepared with relatively nontoxic acids orbases, depending on the particular substituents found on the compoundsdescribed herein. When compounds of the present invention containrelatively acidic functionalities, base addition salts can be obtainedby contacting the neutral form of such compounds with a sufficientamount of the desired base, either neat or in a suitable inert solvent.Examples of pharmaceutically acceptable base addition salts includesodium, potassium, calcium, ammonium, organic amino, or magnesium salt,or a similar salt. When compounds of the present invention containrelatively basic functionalities, acid addition salts can be obtained bycontacting the neutral form of such compounds with a sufficient amountof the desired acid, either neat or in a suitable inert solvent.Examples of pharmaceutically acceptable acid addition salts includethose derived from inorganic acids like hydrochloric, hydrobromic,nitric, carbonic, monohydrogencarbonic, phosphoric,monohydrogenphosphoric, dihydrogenphosphoric, sulfuric,monohydrogensulfuric, hydriodic, or phosphorous acids and the like, aswell as the salts derived from relatively nontoxic organic acids likeacetic, trifluoroacetic, propionic, isobutyric, maleic, malonic,benzoic, succinic, suberic, fumaric, lactic, mandelic, phthalic,benzensulfonic, p-tolylsulfonic, citric, tartaric, methanesulfonic, andthe like. Also included are the salts of amino acids such as arginateand the like, and salts of organic acids like glucuronic or galactunoricacids and the like (see, for example, Berge et al., “PharmaceuticalSalts”, Journal of Pharmaceutical Science, 1977, 66, 1-19). Certainspecific compounds of the present invention contain both basic andacidic functionalities that allow the compounds to be converted intoeither base or acid addition salts.

The neutral forms of the compounds are preferably regenerated bycontacting the salt with a base or acid and isolating the parentcompound in the conventional manner. The parent form of the compounddiffers form the various salt forms in certain physical properties, suchas solubility in polar solvents, but otherwise the salts are equivalentto the parent form of the compound for the purposes of the presentinvention.

The term “low enough pyrogen activity”, with reference to apharmaceutical preparation, refers to a preparation that does notcontain a pyrogen in an amount that would lead to an adverse effect(e.g., irritation, fever, inflammation, diarrhea, respiratory distress,endotoxic shock, etc.) in a subject to which the preparation has beenadministered. For example, the term is meant to encompass preparationsthat are free of, or substantially free of, an endotoxin such as, forexample, a lipopolysaccharide (LPS).

Diseases, Disorders, or Conditions Related to TRPV3 Function

In an embodiment of the methods for preventing or treating a disease ordisorder or condition, the agent being administered is one thatmodulates the level and/or activity of a TRPV3 protein. In certainembodiments, the compound inhibits the expression and/or activity of aTRPV3 protein. In other embodiments, the compound selectively inhibitsthe expression of a TRPV3 protein. In other words, in certainembodiment, the compound inhibits the activity of a TRPV3 proteinpreferentially in comparison to the activity of one or more other ionchannels.

In particular embodiments of the methods for preventing or treatingdiseases and disorders provided herein, the disease or disorder can be,for example, a pain or sensitivity to touch such as pain related to adisease or disorder, e.g., cancer pain, a dermatological disease ordisorder, e.g., psoriasis and basal cell and squamous cell cariconomas,a neurodegenerative disease or disorder, e.g., Alzheimer's disease (AD),Parkinson's disease, Huntington's disease, amyotrophic lateral sclerosis(ALS), and other brain disorders caused by trauma or other insultsincluding aging, an inflammatory disease (e.g., asthma, chronicobstructive pulmonary disease, rheumatoid arthritis, osteoarthritis,inflammatory bowel disease, glomerulonephritis, neuroinflammatorydiseases, multiple sclerosis, and disorders of the immune system),cancer or other proliferative disease, kidney disease and liver disease,a metabolic disorder such as diabetes. Further diseases and conditionsinclude post-surgical pain, post herpetic neuraligia, fibromyalgia, andshingles.

Because of the important role that calcium regulation plays in manycellular processes including cellular activation, gene expression,cellular trafficking and apoptotic cell death, calcium dyshomeostasis isimplicated in the many diseases and disorders involving such cellularactivities. These diseases and disorders include dermatological diseasesand disorders; neurological and neurodegenerative diseases anddisorders; fever associated with various diseases, disorders orconditions; incontinence; inflammatory diseases and disorders such asinflammatory bowel disease and Crohn's disease; respiratory diseases anddisorders such as chronic cough, asthma and chronic obstructivepulmonary disease (COPD); digestive disorders such as ulcers and acidreflux; metabolic diseases and disorders including obesity and diabetes;liver and kidney diseases and disorders; malignancies including cancers;aging-related disorders; and sensitivity to pain and touch.

a. Sensitivity to Pain and Touch, or Pain-Related Diseases or Disorders

Compositions and methods provided herein may be used in connection withprevention or treatment of pain or sensitivity to pain and touch. Painor sensitivity to pain and touch may be indicated in a variety ofdiseases, disorders or conditions, including, but not limited to,diabetic neuropathy, breast pain, psoriasis, eczema, dermatitis, burn,post-herpetic neuralgia (shingles), nociceptive pain, peripheralneuropathic and central neuropathic pain, chronic pain, cancer and tumorpain, spinal cord injury, crush injury and trauma induced pain,migraine, cerebrovascular and vascular pain, Sickle cell disease pain,rheumatoid arthritis pain, musculoskeletal pain including treating signsand symptoms of osteoarthritis and rheumatoid arthritis, orofacial andfacial pain, including dental and cancer related, lower back or pelvicpain, surgical incision related pain, inflammatory and non-inflammatorypain, visceral pain, psychogenic pain and soft tissue inflammatory pain,fibromyalgia-related pain, and reflex sympathetic dystrophy. Thecompounds and methods of the invention may be used in the treatment ofchronic, as well as acute pain. Chronic or acute pain may be the resultof injury, age, or disease.

Other ion channels have been implicated in reception or transmission ofpain. For example, the involvement of N-type calcium channels in thesynaptic transmissions that convey pain signals from sensory afferentnerve cells to the central nervous system has been recognized. Certainnaturally occurring peptide neurotoxins that specifically block N-typecalcium channel have been shown to act as extremely potent and efficientanalgesics in a wide range of animal pain models, including models ofinflammatory and neuropathic pain. The available evidence suggests thatN-type calcium channel blockers are at least as efficacious as opiates,are devoid of a number of the typical opiate side effects (e.g.respiratory depression) and that the analgesic effect is not subject totolerance development.

It has also been shown that potent peripheral analgesia induced by5-alpha-reduced neurosteroid is mediated in part by effects on T-typeCa²⁺ channels (Pathirathna et al., Pain. 2005 April; 114(3):429-43).

Ethosuximide, an anti-epileptic and relatively selective T-type calciumchannel blocker, has also been shown as being highly effective inreversing neuropathic pain caused by the commonly employed cytotoxicspaclitaxel or vincristine (Flatters and Bennett, Pain. 2004 May;109(1-2):150-61).

Pregabalin, a new drug that interacts with the alpha(2)-delta proteinsubunit of the voltage-gated calcium channel, is an efficacious and safetreatment for the pain of diabetic neuropathy (Richter et al., J Pain.2005 April; 6(4):253-60).

The foregoing demonstrate the involvement of various non-TRP channels inthe reception or transmission of pain. Specifically, the foregoingdemonstrate the involvement of various calcium channels in pain.

TRPV3, as well as TRPV1 and TRPV4, are expressed in a pattern consistentwith involvement in pain. TRPV3 is expressed in pain sensitive neurons,and this expression is upregulated following injury (Smith et al.,2002). In addition, TRPV3 is robustly expressed in skin. Accordingly,methods for treating pain include administration of (i) antagonists of aTRPV3 function; (ii) combinations of selective antagonists of a TRPV3and TRPV1 and/or TRPV4 function; or (iii) a pan-TRP inhibitor thatinhibits a function of TRPV3, TRPV1, and TRPV4.

In addition to TRPV family members, other TRP channels have beenimplicated in pain reception and/or sensation. For example, certain TRPMchannels including TRPM8 have been implicated in the reception and/orsensation of pain. Accordingly, in certain embodiments, the methods ofthe present invention include treating pain by administering (i) acombination of a selective TRPV3 antagonist and a selective TRPM8antagonist; (ii) a combination of a selective TRPV3 antagonist, aselective TRPM8 antagonist, and one or more of a selective TRPV1 and/orTRPV4 antagonist; (iii) a cross-TRP inhibitor that antagonizes afunction of TRPV3 and TRPM8; or (iv) a pan inhibitor that antagonizes afunction of TRPV3, TRPM8, and one or more of TRPV1 and TRPV4.

Without being bound by theory, we propose one possible mechanism for howa TRPV3 antagonist may help reduce pain. TRPV3 antagonists can lead tohyperpolarization of the cell. This may lead to a reduction in thefiring of neurons and/or a decrease in action potential frequency. Inaddition, TRPV3 inhibitors may reduce calcium influx into injured cellsand could prevent the calcium dependent changes in gene expression thatsometimes accompany injury. However, regardless of the mechanism ofaction, available expression analysis, electrophysiology andpharmacological efficacy studies support the use of TRPV3 antagonistsfor the treatment of pain.

These findings are somewhat unanticipated because of the uncertainty andcontroversy generated by analysis of TRPV3 knock out mice. It has beenreported that TRPV3 null mice have deficits in their ability to sensetemperature, but not in their ability to sense pain (Moqrich et al.,2005, Science 307: 1468-1472). This finding contradicted an earlierreport that suggested that TRPV3 null mice had normal thermalthresholds, but were unable to develop thermal hyperalgesia in responseto carrageenan or CFA (Smith et al., 2004, Society for NeuroscienceAbstracts).

b. Dermatological Diseases or Disorders

Influx of calcium across plasma membrane of skin cells is a criticalsignaling element involved in cellular differentiation in the skinepidermis (Dotto, 1999 Crit Rev Oral Biol Med 10:442-457). Regulating ormodulating the calcium entry pathway, and thus a critical control pointfor skin cell growth, can treat or prevent skin diseases or disordersthat are characterized by epidermal hyperplasia, a condition in whichskin cells both proliferate too rapidly and differentiate poorly. Suchdiseases include psoriasis, and basal and squamous cell carcinomas.Psoriasis, estimated to affect up to 7 million Americans, afflictssufferers with mild to extreme discomfort, enhanced susceptibility tosecondary infections, and psychological impact due to disfigurement ofthe affected areas (Lebwohl and Ali, 2001 J Am Acad Dermatol45:487-498). Basal cell carcinomas (BCC) and squamous cell carcinomas(SCC) of the skin represent at least one-third of all cancers diagnosedin the United States each year. More than 1 million new cases arereported annually and incidence is increasing. Despite being relativelynon-aggressive, slow-growing cancers, BCCs are capable of significantlocal tissue destruction and disfigurement. SCCs are more aggressive andthus present even greater complications. Further, given that 80% oflesions are on the head and neck with another 15% on shoulders, back orchest, BCCs and SCCs of the skin can have a significant impact on theappearance and quality of life of the afflicted patient.

Many dermatological disorders are accompanied by itch (pruritus).Pruritus and pain share many mechanistic similarities. Both areassociated with activation of C-fibers, both are potentiated byincreases in temperature and inflammatory mediators and both can bequelled with opiates. Decreasing neuronal excitability, particularlyC-fiber excitability may alleviate pruritus associated with dialysis,dermatitis, pregnancy, poison ivy, allergy, dry skin, chemotherapy andeczema.

Acne is a dermatological disorder of complex etiology. Among otherfactors, secretion of oils from the sebaceous glands that contribute tothe development of acne, Since TRPV3 is also expressed in the sebaceousgland and has been shown to be able to regulate secretion in other skincells, antagonizing TRPV3 function might reduce the signs and symptomsof acne.

c. Neurological or Neurodegenerative Diseases and Disorders

Neurodegenerative diseases and disorders include but are not limited toAlzheimer's disease (AD), Parkinson's disease, Huntington's disease,amyotrophic lateral sclerosis (ALS), and other brain disorders caused bytrauma or other insults including aging.

Mechanisms associated with calcium signaling may be altered in manyneurodegenerative diseases and in disorders resulting from brain injury.For example, fibroblasts or T-lymphocytes from patients with AD haveconsistently displayed an increase in Ca²⁺ release from intracellularstores compared to controls (Ito et al. (1994) Proc. Natl. Acad. Sci.U.S.A. 91:534-538; Gibson et al. (1996) Biochem. Biophys. ACTA1316:71-77; Etchenberrigaray et al. (1998) Neurobiology of Disease,5:37-45). Consistent with these observations, mutations in presenilingenes (PS1 or PS2) associated with familial AD (FAD) have been shown toincrease InsP3-mediated Ca²⁺ release from internal stores (Guo et al.(1996) Neuro Report, 8:379-383; Leissring et al. (1999) J.Neurochemistry, 72:1061-1068; Leissring et al. (1999) J. Biol. Chem.274(46):32535-32538; Leissring et al. (2000) J. Biol. Chem.149(4):793-797; Leissring et al. (2000) Proc. Natl. Acad. Sci. U.S.A.97(15):8590-8593). Furthermore, mutations in PS1 or PS2 associated withan increase in amyloidogenic amyloid β peptide generation in AD arereported to be associated with a decrease in intracellular calcium level(Yoo et al. (2000) Neuron, 27(3):561-572).

Experimental traumatic brain injury has been shown to initiate massivedisturbances in Ca²⁺ concentrations in the brain that may contribute tofurther neuronal damage. Intracellular Ca²⁺ may be elevated by manydifferent ion channels. It has been further shown that channel blockersmay be beneficial in the treatment of neurological motor dysfunctionwhen administered in the acute posttraumatic period (Cheney et al.(2000) J. Neurotrauma, 17(1):83-91).

d. Inflammatory Diseases and Disorders

Compositions and methods provided herein may also be used in connectionwith treatment of inflammatory diseases. These diseases include but arenot limited to asthma, chronic obstructive pulmonary disease, rheumatoidarthritis, osteoarthritis, inflammatory bowel disease,glomerulonephritis, neuroinflammatory diseases such as multiplesclerosis, and disorders of the immune system.

The activation of neutrophils (PMN) by inflammatory mediators is partlyachieved by increasing cytosolic calcium concentration ([Ca²⁺]_(i)).Certain calcium channel-mediated calcium influx in particular is thoughtto play an important role in PMN activation. It has been shown thattrauma increases PMN store-operated calcium influx (Hauser et al. (2000)J. Trauma Injury Infection and Critical Care 48 (4):592-598) and thatprolonged elevations of [Ca²⁺]_(i) due to enhanced store-operatedcalcium influx may alter stimulus-response coupling to chemotaxins andcontribute to PMN dysfunction after injury. Modulation of PMN [Ca²⁺]_(i)through store-operated calcium channels might therefore be useful inregulating PMN-mediated inflammation and spare cardiovascular functionafter injury, shock or sepsis (Hauser et al. (2001) J. Leukocyte Biology69 (1):63-68).

Peripheral neuropathy, for example diabetic neuropathy, is a particularcondition that involves both a neuronal and an inflammatory component.Without being bound by a mechanistic theory, the TRPV3 antagonists ofthe invention may be useful in treating peripheral neuropathiesincluding, but not limited to, diabetic neuropathy. In addition to theiruse in the treatment of peripheral neuropathies (e.g., reducinginflammation), the subject inhibitors may also be useful in reducing thepain associated with peripheral neuropathy.

e. Cancer and Other Proliferative Diseases

Compositions and methods provided herein may also be used in connectionwith treatment of malignancies, including, but not limited to,malignancies of lymphoreticular origin, bladder cancer, breast cancer,colon cancer, endometrial cancer, head and neck cancer, lung cancer,melanoma, ovarian cancer, prostate cancer and rectal cancer, in additionto skin cancers described above. Intracellular calcium level may play animportant role in cell proliferation in cancer cells (Weiss et al.(2001) International Journal of Cancer 92 (6):877-882).

In addition, pain associated with cancer or with cancer treatment is asignificant cause of chronic pain. Cancers of the bone, for example,osteosarcoma, are considered exceptionally painful, and patients withadvanced bone cancer may require sedation to tolerate the intense andpersistent pain. Accordingly, TRPV3 antagonists of the inventionrepresent a significant possible therapeutic for the treatment of pain,for example, the pain associated with cancer or with cancer treatment.

Cancer treatments are not only painful, but they may even be toxic tohealthy tissue. Some chemotherapeutic agents can cause painfulneuropathy. Accordingly, TRPV3 antagonists of the invention represent asignificant possible therapeutic for the treatment of the pain and/orinflammation associated with cancer treatments that cause neuropathy.

A major function of prostaglandins is to protect the gastric mucosaIncluded in this function is the modulation of intracellular calciumlevel in human gastric cells which plays a critical role in cellproliferation. Consequently, inhibition of prostaglandins bynonsteroidal anti-inflammatory drugs (NSAIDs) can inhibit calcium influxin gastric cells (Kokoska et al. (1998) Surgery (St Louis) 124(2):429-437). The NSAIDs that relieve inflammation most effectively alsoproduce the greatest gastrointestinal damage (Canadian Family Physician,January 1998, p. 101). Thus, the ability to independently modulatecalcium channels in specific cell types may help to alleviate such sideeffect of anti-inflammatory therapy.

f. Liver Diseases and Disorders

Compositions and methods provided herein may also be used in connectionwith treatment of liver diseases and disorders. These diseases anddisorders include but are not limited to alcoholic liver disease, liverinjury, for example, due to transplantation, hepatitis, cancer, andcirrhosis.

Intracellular calcium level has been implicated in chronic liver disease(Tao et al. (1999) J. Biol. Chem., 274(34):23761-23769) as well astransplantation injury after cold preservation-warm reoxygenation(Elimadi et al. (2001) Am J. Physiology, 281(3 Part 1):G809-G815).Chronic ethanol consumption has been shown to impair liver regeneration,in part, by modulating intracellular calcium level (Zhang et al. (1996)J. Clin. Invest. 98(5): 1237-1244).

g. Kidney Diseases and Disorders

Compositions and methods provided herein may also be used in connectionwith treatment of kidney diseases and disorders. Mesangial cellhyperplasia is often a key feature of such diseases and disorders. Suchdiseases and disorders may be caused by immunological or othermechanisms of injury, including IgAN, membranoproliferativeglomerulonephritis or lupus nephritis. Imbalances in the control ofmesangial cell replication also appear to play a key role in thepathogenesis of progressive renal failure.

The turnover of mesangial cells in normal adult kidney is very low witha renewal rate of less than 1%. A prominent feature of glomerular/kidneydiseases is mesangial hyperplasia due to elevated proliferation rate orreduced cell loss of mesangial cells. When mesangial cell proliferationis induced without cell loss, for example due to mitogenic stimulation,mesangioproliferative glomerulonephritis can result. Data have indicatedthat regulators of mesangial cell growth, particularly growth factors,may act by regulating certain calcium channels (Ma et al. (2001) J. Am.Soc. of Nephrology, 12:(1) 47-53). Modulators of intracellular calciumlevel may aid in the treatment of glomerular diseases by inhibitingmesangial cell proliferation. The epithelial calcium channel CaT2 hasalso been implicated in hypercalciuria and resultant renal stoneformation (Peng et al. (2000) J. Biol. Chem., 275(36):28186-28194).

h. Incontinence

Incontinence is a significant social and medical problem affecting bothmen and women. Incontinence has many causes including, but not limitedto, age, pregnancy, radiation exposure, surgery, injury, and diseases ofthe bladder or musculature that supports the urethra.

Compositions and methods provided herein may be useful in connectionwith the treatment of incontinence. Animal models of incontinence areoften associated with an increase in the frequency of spontaneous actionpotentials and a chronic depolarization of the smooth muscle cells.Evidence suggests that a non-selective cation current could lead to thisdepolarization. Since TRPV3 mRNA is known to be expressed in bladder, aTRPV3 antagonist may be useful in treating incontinence

i. Temperature Regulation

Because of the effects of ion flux on arterial tension and relaxation,the subject compounds can also be used to affect body temperatureregulation, for example, to reduce fever. Furthermore, given that TRPV3channels are heat responsive channels involved in the reception andsensation of heat stimuli, TRPV3 antagonists can be used to modulate thesensation of heat, warmth, or elevated temperatures.

During menopause, many women experience hot flashes. Hot flashes aremarked by sweating, discomfort, and a generally disproportionateexperience of the temperature of one's environment. The symptoms of hotflashes can be very severe, and may interfere with sleep and other dailyactivities. Furthermore, menopause is not only a condition experiencedby women as they age. Premature menopause, and the accompanyingsymptoms, can be induced by hormonal imbalances, certain medications,cancers or other diseases of the female reproductive tract, and partialor total hysterectomies. Thus, menopause and its symptoms may beexperienced by large numbers of women across a diverse age range.

In certain embodiments, TRPV3 antagonists of the present invention canbe used to decrease the perception of heat and temperature associatedwith hot flashes. TRPV3 antagonists of the present invention can beadministered alone, or as part of a therapeutic regimen to decrease thesymptoms associated with menopause. By way of example, TRPV3 antagonistsof the present invention can be administered alone or together withhormone therapy (e.g., estrogen-replacement therapy) used to decreasethe severity of symptoms associated with menopause.

j. Hypertension

Blockers of voltage-gated calcium channels belong to a class ofmedications originally developed to treat hypertension. Such blockersinhibit the movement of calcium into the muscle cells of the heart andarteries. Because calcium is needed for these muscles to contract, suchblockers lower blood pressure by decreasing the force of cardiaccontractile response and relaxing the muscle walls of the arteries.Although TRPV3 is not a voltage-gated calcium channel, it is stillinstrumental in regulating calcium homeostasis, as well as the balanceof other ions, in cells and tissues. Accordingly, TRPV3 antagonists ofthe invention may be used to treat hypertension. Additional uses of thesubject compounds include other conditions that may be ameliorated, inwhole or in part, by relaxing the muscle walls of blood vessels.Exemplary conditions include headaches and migraine attacks.

k. Hair Loss

TRPV3 knock-out mice have a significant hair loss phenotype.Accordingly, the TRPV3 antagonists of the present invention can be usedto promote hair loss or to otherwise inhibit the growth of body hair. Byway of example, TRPV3 antagonists can be applied topically to the skinto promote the loss of or to otherwise inhibit the growth of body hair.In such embodiments, the TRPV3 antagonists act as a depilatory agent topromote the loss of or to otherwise inhibit the growth body hair. Whenused in this manner, one or more TRPV3 antagonist can be used alone orin combination with an additional depilatory agent. Additionally, one ormore TRPV3 antagonist can be used to supplement other hair removaltechniques such as waxing or shaving. In such a way, a TRPV3 antagonistcan be used alone or as part of a hair removal regimen to reduce oreliminate unwanted body hair. Exemplary unwanted body hair includes, butis not limited to, hair on the legs, arms, back, upper lip, chest,bikini area, underarms, and buttocks.

Additionally or alternatively, TRPV3 antagonists can be administeredsystemically to promote the loss of or to prevent the growth of bodyhair.

In any of the foregoing, TRPV3 antagonists likely provide an improvedmethod for reducing or eliminating unwanted body hair. Given the paininhibiting activity of TRPV3 antagonists, their use alone or as part ofa hair removal regimen provides an improved method for removing bodyhair with less discomfort than currently available waxes and chemicaldepilatories.

As outlined above, compounds that antagonize a function of TRPV3 can beused in the treatment of many diseases, injuries, disorders, andconditions. In certain embodiments, TRPV3 inhibitors can be used in thetreatment of pain. As outlined above, TRPV3 inhibitors can be used inthe treatment of pain resulting from injury or disease, as well as painexperienced as a consequence of treatment. Exemplary classes of paininclude nociceptive pain, inflammatory pain, and neuropathic pain. Suchpain can be chronic or acute. TRPV3 inhibitors can be used in thetreatment of one or more of any of the foregoing classes of pain. Incertain embodiments, TRPV3 inhibitors can be used in the treatment ofnociceptive pain. In certain other embodiments, TRPV3 inhibitors can beused in the treatment of inflammatory pain. In certain otherembodiments, TRPV3 inhibitors can be used in the treatment ofneuropathic pain.

As outlined above, TRPV3 inhibitors may be particularly useful in thetreatment of pain associated with cancer, osteoarthritis, rheumatoidarthritis, post-herpetic neuralgia, burns, and other indicationsdetailed above. To further illustrate, additional exemplary indicationsfor which compounds of the present invention can be used include oralpain, Fabry's disease, and fibromyalgia syndrome.

Fabry's Disease

Vague complaints of pain in hands and feet may be a presenting feature.These symptoms are called acroparesthesias, as they reflect theperipheral neuropathy that is a frequent manifestation of the disease.This pain may be both episodic and chronic. Acute episodes may betriggered by exposure to extremes of temperature, stress, emotion,and/or fatigue.

Fibromyalgia

Fibromyalgia (FMS; fibromyalgia syndrome) is a widespreadmusculoskeletal pain and fatigue disorder for which the cause is stillunknown. Fibromyalgia is characterized by pain in the muscles,ligaments, and tendons. Most patients with FMS say that they ache allover. The condition affects more women than men, and occurs in people ofall ages. Overall, FMS is estimated to afflict 3-6% of the population.

Patients have described the pain associated with fibromylagia as deepmuscular aching, throbbing, shooting, and stabbing. The pain sometimesincludes an intense burning sensation. The pain and stiffness are oftenworse in the morning or after repetitive use of a particular musclegroup.

In addition to pain, varying levels of fatigue are often associated withfibromylagia. Some patients report only mild fatigue while for othersthe fatigue is incapacitating. Other symptoms of fibromylagia includegastrointestinal symptoms. Irritable bowel syndrome and IBS-likesymptoms such as constipation, diarrhea, frequent abdominal pain,abdominal gas, and nausea occur in roughly 40 to 70% of FMS patients.Acid reflux or gastroesophogeal reflux disease (GERD) occurs at asimilar frequency.

Another frequent and debilitating symptom of FMS is chronic headaches,including migraine and tension-type headaches. Such headaches areexperienced by approximately 70% of FMS patients. Additionally, FMSpatients often experience temporomandibular joint dysfunction syndrome(also known as TMJ). TMJ produces pain in the jaw, teeth, and mouth, andmay also exacerbate headaches.

Other common symptoms of FMS include, but are not limited to,premenstrual syndrome and painful periods; chest pain; morningstiffness; cognitive or memory impairment; numbness and tinglingsensations; muscle twitching; irritable bladder; the feeling of swollenextremities; skin sensitivities; dry eyes and mouth; dizziness; andimpaired coordination. Additionally, patients are often sensitive toodors, loud noises, and bright lights.

The cause of FMS remains unknown. However, the onset of the disorder hasbeen linked to infections (viral or bacterial), rheumatoid arthritis,lupus, and hypothyroidism. The link between these and other possibletriggers is unclear.

The impact of FMS on the patient is directly correlated with the levelof pain and fatigue. Some patients experience pain so severe as tointerfere with normal work or family functioning. There is currently nocure for FMS, and current therapies focus primarily on improving sleep(to decrease fatigue) and treating pain. Compounds of the presentinvention could be used to help manage the pain associated with FMS.Such pain includes, but is not limited to, oral pain in the jaw, teeth,and mouth. Such pain also includes non-oral musco-skeletal pain, paindue to headaches, and pain due to gastrointestinal symptoms.

Oral pain is a particular category of pain that may be treated using theTRPV3 inhibitors of the present invention. The term “oral pain” refersto any pain in the mouth, throat, lips, gums, teeth, tongue, or jaw. Theterm is used regardless of the cause of the pain and regardless ofwhether the oral pain is a primary or secondary symptom of a particulardisease, injury, or condition.

Oral pain has a large number of possible causes. In certain embodiments,oral pain is caused by an injury or disease of the mouth, jaw, teeth,gums, throat, lips, or tongue. In certain other embodiments, oral painis a consequence of an injury or disease that primarily affects anotherpart of the body. In still other embodiments, oral pain is a side effectof a therapy used to treat an injury or disease of the mouth or anotherpart of the body. TRPV3 inhibitors are useful in treating oral painregardless of its cause.

All pain has a serious negative impact on the health and well being ofthe sufferer. However, oral pain may have a particularly deleteriousimpact on patient health and quality of life. In particular, oral paincan interfere with appropriate eating and drinking. Thus, individualswith oral pain are susceptible to weight loss, malnutrition, anddehydration. In some instances, oral pain may interfere with hydrationand nutrition so significantly as to require intravenous, nasogastric,or other artificial support (e.g., tube feeding and/or hydration).Additionally, oral pain can interfere with proper oral hygiene. Poororal hygiene may further exacerbate many of the causes of oral pain, forexample, oral pain due to infection or abscess.

In certain embodiments, oral pain is caused by ulcers, sores, or otherlesions in the mouth. For example, oral pain may be caused by ulcers,sores, or other lesions on the tongue, gums, lips, throat, or othertissues of the mouth. Alternatively or additionally, oral pain may becaused by inflammation of the throat, tongue, gums, lips, or othertissues of the mouth. Inflammation may accompany ulcers or otherlesions, or inflammation may occur prior to or in the absence offormation of ulcers or other lesions.

The invention contemplates treatment of oral pain by administering aTRPV3 inhibitor by any route of administration described herein. Incertain embodiments, TRPV3 inhibitors for use in the treatment of oralpain are administered orally. Preferred preparations for oraladministration of TRPV3 inhibitors for use in treating oral pain are asa mouthwash, a gel, a tooth paste or other paste, a liquid, a lozenge,via a swab, or in association with a mouth guard or dental apparatus.The preparation and particular method of administration will depend onthe cause of the oral pain, the overall health and underlying medicalconditions of the patient, the severity of the pain, and othermedications or therapies the patient is concurrently receiving. Amedical practitioner can readily determine the optimal formulation foruse in a particular patient.

The conditions provided below are intended to illustrate the range ofinjuries and diseases of diverse etiology that may lead to oral pain.The invention contemplates administration of a TRPV3 inhibitor,according to the present invention, to treat or prevent oral pain. Incertain embodiments, compounds of the invention can be orallyadministered, for example as a gel, paste, mouth wash, or other oralpreparation, to help treat or prevent oral pain associated with anyinjury, disease, or condition. Regardless of the particular formulation,the invention contemplates administration by, for example, directapplication to the affected area of the mouth, rinsing of the entiremouth, via a swab, via a syringe, or on a mouth guard or other dentalapparatus.

For any of these conditions, the invention contemplates administrationof a TRPV3 inhibitor alone, or in combination with one or more othercompounds or treatment regimens appropriate for the particular injury orcondition.

Oral Mucositis

Oral mucositis, also known as stomatitis, is a common complication ofmany cancer treatments. Patients receiving systemic chemotherapy and/orlocal radiotherapy often develop extremely painful ulcers of the oralmucosa. This side effect is not limited to patients suffering fromcancers of the head and neck, but rather is a debilitating side effectafflicting approximately 40% of all chemotherapy patients (Preventionand Treatment of Oral Mucositis in Cancer Patients, 1998, Best Practice:2, pages 1-6.)

Oral mucositis is extremely painful. Additionally, oral mucositisinterferes with proper nutrition and hydration of cancer patients. Giventhe already compromised status of patients undergoing chemotherapyand/or radiotherapy, further interference with nutrition and hydrationmay seriously undermine patient health. Furthermore, these ulcerspresent an increased risk of infection. This risk is particularly acutein patients with compromised immune systems. Examples of patients atparticular risk of developing an opportunistic infection are patientswhose treatment included removal of one or more lymph nodes, patientswho previously received high-dose chemotherapy in preparation for a bonemarrow or stem cell transplant, and patients with an underlyingimmunosuppressive disorder (e.g., HIV or hepatitis).

Canker Sores

Canker sores, also known as aphthous ulcers (aphthae), may be relativelysmall and out-of-sight. However, they are often painful, persistent andannoying. Canker sores are shallow ulcers in the mouth that can makeeating and talking uncomfortable. They may occur on the tongue, softpalate, inside the cheek or lip, or at the base of the gums. Cankersores differ from cold sores in that they occur on the internal softtissues of the mouth and aren't contagious. Conversely, cold soresalmost always start out on the lips and don't often spread to the softtissues of the mouth. In addition, cold sores are caused by a form ofthe herpes virus, making them extremely contagious.

The cause of most canker sores remains a mystery. Researchers generallybelieve that stress or tissue injury may cause the eruption of cankersores. In some cases a minor injury, for example biting the inside ofthe mouth or eating rough foods, may trigger a canker sore. Other causesmay include: (i) faulty immune system function; (ii) nutritionalproblems, such as a deficiency of vitamin B-12, zinc, folic acid oriron; (iii) diseases of the gastrointestinal tract; (iv) food allergies;or (v) the menstrual cycle.

Canker sores are common, but the cause in many cases is unknown. Cankersores can occur at any age, but often they first appear between the agesof 10 and 40 years. Although canker sores typically resolve on theirown, they can be very uncomfortable.

Dental/Tooth Abscess

Infection or decay can lead to an abscess. An abscess may have seriousdental and medical consequences. For example, a severe infection causedby a dental abscess may lead to a sinus or systemic infection.Furthermore, an abscess may lead to the need to extract one or moreteeth. Extraction may be necessary due to significant tooth decay, orbecause the infection is too severe to fully treat in the presence ofthe offending tooth.

Regardless of the ultimate outcome, a dental abscess may be extremelypainful. Not only is the pain uncomfortable, but it may interfere withproper nutrition and hydration. Methods and compositions for reducingthe pain associated with dental abscess would provide significantbenefits for their management.

Gastroesophageal Reflux Disease

Gastroesophageal reflux disease, or GERD, occurs when the loweresophageal sphincter (LES) does not close properly and stomach contentsleak back into the esophagus. The LES is a ring of muscle at the bottomof the esophagus that acts like a valve between the esophagus andstomach. When refluxed stomach acid touches the lining of the esophagus,it causes a burning sensation in the chest or throat. This is oftenexperienced as heartburn. The refluxed fluid may even be tasted in theback of the mouth, a sensation commonly referred to as acid indigestion.

Although occasional heartburn is uncommon and not necessarily indicativeof GERD, heartburn that occurs more than twice a week may be a sign ofGERD. In addition to the discomfort of heartburn and indigestion, GERDmay lead to other serious health problems. For example, over time, acidrefluxed to the back of the throat can lead to oral sores, lesions, orulcers in the mouth, gums, tongue, throat, or lips. The lesions cancause significant pain, can interfere with nutrition and hydration, andcan leave a person vulnerable to infection.

Administration of TRPV3 inhibitors, according to the present invention,may be useful in treating oral pain from lesions caused by GERD. TRPV3inhibitors may be used as part of a treatment regimen where the TRPV3inhibitor is administered to help manage the discomfort of the orallesion, while other agents or therapeutics interventions are used tomanage the GERD.

Gingivostomatitis

Gingivostomatitis is a disorder involving sores on the mouth and gumsthat result from a viral infection. Gingivostomatitis is characterizedby inflammation of the gums and mucosa and multiple oral ulcers. Theinflammation and ulcers are caused by viral infections, particularlythose that cause common childhood illness such as herpes virus (coldsores and acute herpetic stomatitis), and Coxsackie viruses (hand, footand mouth disease and herpangina). These viruses cause shallow ulcerswith a grayish or yellowish base and a slightly red margin, on thetissues of the gums (gingiva), the lining of the cheeks (buccal mucosa),or other soft tissues of the mouth. Although this condition can occur inpatients of any age, it is particularly common in children.

The oral ulcers casued by these viruses can be very painful. The ulcersare often accompanied by a fever. Overall, the condition can takeseveral weeks to resolve. The recognized treatments forgingivostomatitis focus on reducing the pain caused by the oral ulcers.This is particularly important for children who may refuse food orliquids because of their discomfort, thus making them especiallysusceptible to dehydration.

Oral Thrush

Oral thrush is a fungal infection generally caused by the yeast fungus,Candida albicans, in the mucous membranes of the mouth. Strictlyspeaking, thrush is only a temporary Candida infection in the oralcavity of babies. However, the term is used generally to refer to fungalinfections in the mouths and throats of children and adults.

Candida is present in the oral cavity of almost half of the population.For example, everyone who wears dentures has Candida, withoutnecessarily suffering any ill effects. Generally, Candida does notcreate problems until there is a change in the chemistry of the oralcavity such that the growth of Candida is favored over the othermicroorganisms that typically inhabit the mouth and throat. Changes inoral chemistry sufficient to permit the growth of Candida may occur as aside effect to taking antibiotics or chemotherapeutics. Overall patienthealth may also influence the chemistry of the mouth. HIV infection,diabetes, malnutrition, age, and immunodeficiency are exemplaryconditions that can shift oral chemistry enough to permit the overgrowthof Candida in the mouth and throat.

In addition to shifts in oral chemistry, people whose dentures don't fitwell can sustain breaks in the mucous membranes in their mouth. Thesebreaks provide an opportunity for Candida infection in the mouth andlips.

Thrush causes white, cream-colored, or yellow spots in the mouth. Thespots are slightly raised. If these spots are scraped they tend tobleed. Thrush can be very uncomfortable, and may cause a burningsensation in the mouth and throat. The discomfort may interfere withhydration and nutrition. Furthermore, the discomfort may interfere withproper oral hygiene such as brushing and flossing.

Standard treatment of thrush is by administration of anti-fungal agents.These agents can be administered directly to the mouth, for example, inthe form of pastilles that are sucked or oral suspensions that are heldin the mouth before swallowing. Examples include nystatin (e.g., Nystanoral suspension), amphotericin (e.g., Fungilin lozenges) or miconazole(e.g., Daktarin oral gel). In addition to standard anti-fungal therapy,compounds of the present invention can be administered to manage thepain and discomfort associated with thrush.

Glossitis

Glossitis is an abnormality of the tongue that results frominflammation. Glossitis occurs when there is acute or chronicinflammation of the tongue. It causes the tongue to swell and changecolor. Finger-like projections on the surface of the tongue (papillae)are lost, causing the tongue to appear smooth. Glossitis has a number ofcauses including, but not limited to, the following: bacterialinfections; viral infections (including oral herpes simplex); injury ortrauma; exposure to irritants (e.g., tobacco, alcohol, hot foods,spices); allergic reactions; vitamin or mineral deficiencies (e.g., irondeficiency anemia, pernicious anemia and other B-vitamin deficiencies);or as a side effect of other diseases or disorders.

The symptoms of glossitis include swelling, soreness, and tenderness ofthe tongue. Additionally, the tongue often changes appearance, becomingsmooth and dark red in color. As a consequence of the swelling anddiscomfort, glossitis often makes chewing, swallowing, and speakingdiffcult.

The typical treatment for glossitis depends on the underlying cause ofthe inflammation. Regardless of the particular antibiotics,anti-inflammatories, or anti-viral agents that may be administered tocombat the underlying cause of glossitis, compounds according to thepresent invention may be administered to decrease the pain anddiscomfort associated with glossitis. Decreasing the pain associatedwith glossitis is especially important when it interferes with propernutrition and hydration, or when it interferes with or prevents properoral hygiene.

Cutaneous Diseases

Oral ulcers may result from any of a number of cutaneous diseases. Forexample, lichen planus, pemphigus, pemphigoid, and erythema multiformemay lead to oral ulcers. Such oral ulcers may cause significant painthat can be treated using the compounds of the present invention.

Reduction of pain may help facilitate healing. This is especiallyimportant for patients with pemphigus and pemphigoid who develop oralulcers. Such patients are already immunosuppressed, and may thus be moresusceptible to opportunistic infections from lesions in the mouth.

Gastrointestinal Diseases

Oral ulcers may result from any of a number of gastrointestinaldiseases. Conditions which interfere with proper digestion, managementand flow of stomach and other digestive acids, motility, and eliminationmay lead to oral ulcers and other lesions. In some instances, the oralulcers are the results of acids or partially digested food refluxinginto the esophagus. In other instances, the oral ulcers result fromfrequent vomiting. In still other instances, oral ulcers occur due tovitamin deficiency, mineral deficiency or other nutritional deficiencysecondary to the gastrointestinal disease. In still other instances,oral ulcers are part of the complex etiology that characterizes thegastrointestinal disease.

Oral ulcers resulting from or experienced as part of a gastrointestinaldisease may be extremely painful. They may undermine proper nutritionand hydration for a patient whose underlying gastrointestinal diseasemay already impose multiple limitations on diet. Accordingly, methodsand compositions for decreasing the discomfort and pain associated withthese oral ulcers offer substantial benefits for patients with anunderlying gastrointestinal condition.

Exemplary gastrointestinal conditions which may lead to oralinflammation, lesions, or ulcers include, but are not limited to,Crohn's disease, ulcerative colitis, irritable bowel syndrome, celiacsprue, and dermatitis herpetiformis. The primary symptoms of theseconditions may be managed with diet, stress management, and medications.The TRPV3 inhibitors of the present invention may be used to help managethe pain and discomfort of oral inflammation, lesions, or ulcers causedby any of these gastrointestinal conditions.

Rheumatoid Diseases

A consequence of several rheumatoid diseases is oral ulcers. Forexample, lupus, Behcet's syndrome, Sweet's syndrome, and Reiter'sdisease may all lead to oral ulcers. Such oral ulcers may causesignificant mouth pain that can be treated using the compounds of thepresent invention.

Sjogren's Syndrome

Dry mouth is a common symptom associated with Sjögren's syndrome. Drymouth is caused by a decrease in the production of saliva. Saliva is anessential body fluid for protection and preservation of the oral cavityand oral functions. Although saliva is mostly water, it also containsover 60 substances which serve the following important functions:protect, lubricate and cleanse the oral mucosa; aid chewing, swallowingand talking; protect the teeth against decay; protect the mouth, teeth,and throat from infection by bacteria, yeasts, and viruses; support andfacilitate our sense of taste.

Given the important functions of saliva, decreased salivation can leadto many problems. If the condition persists for months or years, apatient may develop oral complications such as difficulty swallowing,severe and progressive tooth decay, oral infections (particularlyfungal), or combinations of these. Many of the conditions can causediscomfort, in their own right, and may also lead to oral lesions orulcers.

Several medications are available to help increase salivary secretion inpatients with dry mouth. Pilocarpine (Salagen®) and cevimeline (Evoxac®)reduce symptoms of dry mouth and increase salivary secretion. However,these drugs don't prevent tooth decay or treat the oral pain associatedwith the symptoms or effects of dry mouth.

Vitamin or Mineral Deficiencies

In some instances, vitamin or mineral deficiencies may lead to ulcers orother sores in the mouth. For example, deficiency in vitamin C may leadto the oral lesions characteristic of scurvy. Deficiencies in vitaminsB1, B2, B6, or B12 may also lead to oral lesions. Additionally,deficiencies in zinc, folic acid, iron, selenium, or calcium may lead tooral lesions.

In certain embodiments, a vitamin or mineral deficiency is aprecipitating factor leading to a canker sore. However, a vitamin ormineral deficiency may also lead to other types of oral ulcers andlesions. Regardless of the nature of the lesion, compounds of thepresent invention can be used to help manage the associated pain.

Allergies

Allergies can sometimes lead to canker sores and other oral lesions.Oral lesions due to an allergy may be more likely when a person's oraltissues come into contact with the causative allergen. However, contactbetween the allergen and oral tissue is not necessarily required toproduce an oral lesion. Exemplary allergens that can lead to orallesions include food allergens such as fruits and vegetables (e.g.,strawberries, lemons, oranges, pineapples, apples, figs, tomatoes);shellfish; chocolate; nuts; dairy (e.g., milk and cheese); cereal grains(e.g., buckwheat, wheat, oats, rye, barley, gluten protein found ingrains); additives (e.g., cinnamonaldehyde (a flavoring agent), benzoicacid (a preservative); toothpastes (e.g., some people have a sensitivityto sodium laurel sulfate found in certain toothpastes and mouthwashes);nonsteroidal anti-inflammatory drugs (NSAIDs; some people have asensitivity leading to canker sores in response to this class of drug).

Other Exemplary Conditions and Injuries

The foregoing are merely exemplary of diseases and conditions that causeor lead to inflammation, lesions, ulcers, or other sources of oral pain.In other embodiments, the oral pain is due to an injury to the mouth,jaw, lips, gums, or teeth. In other embodiments, the oral pain is due tooral surgery, for example, surgery for cancer, tooth extraction, or jawremodeling. Other conditions that may lead to oral ulcers, and thus oralpain, include, but are not limited to chickpox, herpes zoster,infectious mononucleosis, syphilis, tuberculosis, acute necrotizinggingivitis, and burning mouth syndrome. Additionally, conditions thatlead to a compromised immune system put patients at risk for, amongother complications, oral inflammation, lesions, or ulcers. HIVinfection, AIDS, and hepatitis are all conditions that undermine theimmune system and may lead to oral lesions or ulcers. Additionally,individuals taking immunosuppressants (e.g., organ transplantrecipients, bone marrow recipients, stem cells recipients, patients withan autoimmune disease) are at increased risk of developing painful orallesions.

The invention contemplates the use of TRPV3 inhibitors, according to thepresent invention, in the treatment of oral pain regardless of theunderlying cause. In certain embodiments, TRPV3 inhibitors for treatingoral pain can be administered orally, for example, as a paste, mouthwash, gel, or other liquid preparation. In certain embodiments, thepaste, mouth wash, gel, or other liquid preparation is administered viaa swab, mouth guard, or other dental apparatus. In certain embodiments,the preparation is applied locally to the mouth, but is not otherwiseingested. For example, a mouth wash formulation that is not swallowedmay be used. Regardless of the formulaion and route of administeration,the invention contemplates administration of the subject TRPV3inhibitors as part of an overall treatment strategy that also includestherapies appropriate for the particular disease or condition thatcaused the oral inflammation, lesion, or ulcer.

TRPV3 inhibitors may be used to treat oral pain resulting from any ofthe foregoing injuries, diseases, or conditions. Additionally,Applicants note that the subject TRPV3 inhibitors may also be useful inthe treatment of the underlying aforementioned diseases and conditionsthemselves. Specifically, TRPV3 inhibitors may be useful in thetreatment of inflammation, and thus diseases or conditions with aninflammatory component, whether the symptoms manifest themselves in themouth or in other parts of the body, may themselves be treatable withthe subject TRPV3 inhibitors. Accordingly, the invention contemplatesand recognizes that for some conditions the therapeutic affects ofadministering a TRPV3 inhibitor may be two-fold: (i) decreasing painassociated with one or more symptoms of a disease or condition and (ii)treating the underlying symptoms or disease.

Disease and Injury Models

Compounds that antagonize TRPV3 function may be useful in theprophylaxis and treatment of any of the foregoing injuries, diseases,disorders, or conditions. In addition to in vitro assays of the activityof these compounds, their efficacy can be readily tested in one or moreanimal models. By way of example, numerous well known animal modelsexist. One or more suitable animal models (e.g., suitable in light ofthe particular indication) can be selected.

Pain can be generally categorized as chronic pain and acute pain. Thetwo categories of pain differ in duration, as well as underlyingmechanism. Chronic pain is not only persistent, but also does notgenerally respond well to treatment with currently available analgesics,non-steroidal anti-inflammatory drugs, and opioids.

Two broad sub-categories of chronic pain are neuropathic pain and cancerpain. Wang and Wang (2003) Advanced Drug Delivery Reviews 55: 949-965.Neuropathic pain refers to pain resulting from damage (e.g., fromdisease, injury, age) to the nervous system (e.g., nerves, spinal cord,CNS, PNS). Cancer-related pain may be caused by tumor infiltration,nerve compression, substances secreted by tumors, or the particulartreatment regimen (e.g., radiation, chemotherapeutics, surgery).

Pain is also often classified mechanistically as nociceptive,inflammatory, or neuropathic. Nociceptive pain is pain experiencedfollowing, for example, changes or extremes in temperature, exposure toacids, exposure to chemical agents, exposure to force, and exposure topressure. Reception of painful stimuli sends impulses to the dorsal rootganglia. The response is typically a combination of a reflexive response(e.g., withdrawal from the stimuli) and an emotional reaction.Inflammation is the immune system's response to injury or disease. Inresponse to injury or disease, macrophages, mast cells, neutrophils, andother cells of the immune system are recruited. This infiltration ofcells, along with the release of cytokines and other factors (e.g.,histamine, serotonin, bradykinin, prostaglandins, ATP, H+, nerve growthfactor, TNFα, endothelins, interleukins), can cause fever, swelling, andpain. Current treatments for the pain of inflammation include Cox2inhibitors and opioids. Neuropathic pain refers to pain resulting fromdamage (e.g., from disease, injury, age) to the nervous system (e.g.,nerves, spinal cord, CNS, PNS). Current treatment for neuropathic painincludes tricyclic antidepressants, anticonvulsants, Na+ channelblockers, NMDA receptor antagonists, and opioids.

There are numerous animal models for studying pain. Generally, theanimal models mimic one of the foregoing mechanisms of pain, rather thanthe pain associated with any one disease or injury. Such models provideevidence of whether a drug or therapy would be effective in treating anyof a number of injuries, diseases, or conditions that generate pain viaa particular mechanism (e.g., nociceptive, inflammatory, orneuropathic).

The various models use various agents or procedures to simulate painresulting from injuries, diseases, or other conditions. Blackburn-Munro(2004) Trends in Pharmacological Sciences 25: 299-305 (see, for example,Table 1). Behavioral characteristics of challenged animals can then beobserved. Compounds or procedures that may reduce pain in the animalscan be readily tested by observing behavioral characteristics ofchallenged animals in the presence versus the absence of the testcompound(s) or procedure.

Exemplary behavioral tests used to study chronic pain include tests ofspontaneous pain, allodynia, and hyperalgesia. Id. To assess spontaneouspain, posture, gait, nocifensive signs (e.g., paw licking, excessivegrooming, excessive exploratory behavior, guarding of the injured bodypart, and self-mutilation) can be observed. To measure evoked pain,behavioral responses can be examined following exposure to heat (e.g.,thermal injury model).

Exemplary animal models of pain include, but are not limited to, theChung model, the carageenan induced hyperalgesia model, the Freund'scomplete adjuvant (CFA) induced hyperalgesia model, the thermal injurymodel, the formalin model and the Bennett Model. The Chung model ofneuropathic pain (without inflammation) involves ligating one or morespinal nerves. Chung et al. (2004) Methods Mol Med 99: 35-45; Kim andChung (1992) Pain 50: 355-363. Ligation of the spinal nerves results ina variety of behavioral changes in the animals including heathyperalgesia, cold allodynia, and ongoing pain. Compounds thatantagonize TRPV3 can be administered to ligated animals to assesswhether they diminish these ligation induced behavioral changes incomparison to that observed in the absence of compound.

Carageenan induced hyperalgesia and Freund's complete adjuvent (CFA)induced hyperalgesia are models of inflammatory pain. Walker et al.(2003) Journal of Pharmacol Exp Ther 304: 56-62; McGaraughty et al.(2003) Br J Pharmacol 140: 1381-1388; Honore et al. (2005) J PharmacolExp Ther. Compounds that antagonize TRPV3 can be administered tocarrageenan or FCA challenged animals to assess whether they diminishthermal hyperalgesia in comparison to that observed in the absence ofcompound. In addition, the ability of compounds that antagonize TRPV3function to diminish cold and/or mechanical hypersensitivity can also beassessed in these models. Typically, the carrageenan inducedhyperalgesia model is believed to mimic acute inflammatory pain and theCFA model is believed to mimic chronic pain and chronic inflammatorypain.

The Bennett model uses prolonged ischemia of the paw to mirror chronicpain. Xanthos et al. (2004) J Pain 5: S1. This provides an animal modelfor chronic pain including post-operative pain, complex regional painsyndrome, and reflex sympathetic dystrophy. Prolonged ischemia inducesbehavioral changes in the animals including hyperalgesia to mechanicalstimuli, sensitivity to cold, pain behaviors (e.g., paw shaking,licking, and/or favoring), and hyperpathia. Compounds that antagonizeTRPV3 can be administered to challenged animals to assess whether theydiminish any or all of these behaviors in comparison to that observed inthe absence of compound. Similar experiments can be conducted in athermal injury model which can be used to mimic post-operative pain.

Additional models of neuropathic pain include central pain models basedon spinal cord injury. Chronic pain is generated by inducing a spinalcord injury, for example, by dropping a weight on a surgically exposedarea of spinal cord (e.g., weight-drop model). Spinal cord injury canadditionally be induced by crushing or compressing the spinal cord, bydelivering neurotoxin, using photochemicals, or by hemisecting thespinal cord. Wang and Wang (2003).

Additional models of neuropathic pain include peripheral nerve injurymodels. The term peripheral neuropathy encompasses a variety ofdiseases, conditions, and injuries. One of skill in the art can readilyselect an appropriate model in light of the particular condition ordisease under investigation. Exemplary models include, but are notlimited to, the neuroma model, the Bennett model, the Seltzer model, theChung model (ligation at either L5 or L5/L6), the sciatic cryoneurolysismodel, the inferior caudal trunk resection model, and the sciaticinflammatory neuritis model. Id.

Exemplary models of neuropathic pain associated with particular diseasesare also available. Diabetes and shingles are two diseases oftenaccompanied by neuropathic pain. Even following an acute shinglesepisodes, some patients continue to suffer from postherpetic neuralgiaand experience persistent pain lasting years. Neuropathic pain caused byshingles and/or postherpetic neuralgia can be studied in thepostherpetic neuralgia model (PHN). Diabetic neuropathy can be studiedin diabetic mouse models, as well as chemically induced models ofdiabetic neuropathy. Wang and Wang (2003).

As outlined above, cancer pain may have any of a number of causes, andnumerous animal models exist to examine cancer pain related to, forexample, chemotherapeutics or tumor infiltration. Exemplary models oftoxin-related cancer pain include the vincristine-induced peripheralneuropathy model, the taxol-induced peripheral neuropathy model, and thecisplatin-induced peripheral neuropathy model. Wang and Wang (2003). Anexemplary model of cancer pain caused by tumor infiltration is thecancer invasion pain model (CIP). Id.

Primary and metastatic bone cancers are associated with tremendous pain.Several models of bone cancer pain exist including the mouse femur bonecancer pain model (FBC), the mouse calcaneus bone cancer pain model(CBC), and the rat tibia bone cancer model (TBC). Id.

An additional model of pain is the formalin model. Like the carrageenanand CFA models, the formalin model involves injection of an irritantintradermally or intraperitoneally into an animal. Injection offormalin, a 37 percent solution of formaldehyde, is the most commonlyused agent for intradermal paw injection (the formalin test). Injectionof a 0.5 to 15 percent solution of formalin (usually about 3.5%) intothe dorsal or plantar surface of the fore- or hindpaw produces abiphasic painful response of increasing and decreasing intensity forabout 60 minutes after the injection. Typical responses include the pawbeing lifted, licked, nibbled, or shaken. These responses are considerednociceptive. The initial phase of the response (also known as the EarlyPhase), which lasts 3 to 5 minutes, is probably due to direct chemicalstimulation of nociceptors. This is followed by 10 to 15 minutes duringwhich animals display little behavior suggestive of nociception. Thesecond phase of this response (also known as the Late Phase) startsabout 15 to 20 minutes after the formalin injection and lasts 20 to 40minutes, initially rising with both number and frequency of nociceptivebehaviors, reaching a peak, then falling off. The intensities of thesenociceptive behaviors are dependent on the concentration of formalinused The second phase involves a period of sensitization during whichinflammatory phenomena occur. The two phases of responsiveness toformalin injection makes the formalin model an appropriate model forstudying mociceptive and acute inflammatory pain. It may also model, insome respects, neuropathic pain.

In addition to any of the foregoing models of chronic pain, compoundsthat antagonize TRPV3 function can be tested in one or more models ofacute pain. Valenzano et al. (2005) Neuropharmacology 48: 658-672.Regardless of whether compounds are tested in models of chronic pain,acute pain, or both, these studies are typically (though notexclusively) conducted, for example, in mice, rats, or guinea pigs.Additionally, compounds can be tested in various cell lines that providein vitro assays of pain. Wang and Wang (2003).

The foregoing animal models are relied upon extensively in the study ofpain. The following provide additional exemplary references describingthe use of these models in the study of pain: thermal injury model(Jones and Sorkin, 1998, Brain Res 810: 93-99; Nozaki-Taguchi and Yaksh,1998, Neuroscience Lett 254: 25-28; Jun and Yaksh, 1998, Anesth Analg86: 348-354), formalin model (Yaksh et al., 2001, J Appl Physiol 90:2386-2402), carrageenan model (Hargreaves et al., 1988, Pain 32: 77-88),and CFA model (Nagakura et al., 2003, J Pharmacol Exp Ther 306:490-497).

For testing the efficacy of TRPV3 antagonists for the treatment ofcough, experiments using the conscious guinea pig model of cough can bereadily conducted. Tanaka and Maruyama (2003) Journal Pharmacol Sci 93:465-470; McLeod et al. (2001) Br J Pharmacol 132: 1175-1178. Briefly,guinea pigs serve as a useful animal model for cough because, unlikeother rodents such as mice and rats, guinea pigs actually cough.Furthermore, guinea pig coughing appears to mimic human coughing interms of the posture, behavior, and appearance of the coughing animal.

To induce cough, conscious guinea pigs are exposed to an inducing agentsuch as citric acid or capsaicin. The response of the animal is measuredby counting the number of coughs. The effectiveness of a coughsuppressing agent, for example a compound that inhibits TRPV3, can bemeasured by administering the agent and assessing the ability of theagent to decrease the number of coughs elicited by exposure to citricacid, capsaicin, or other similar cough-inducing agent. In this way,TRPV3 inhibitors for use in the treatment of cough can be readilyevaluated and identified.

Additional models of cough include the unconscious guinea pig model.Rouget et al. (2004) Br J Pharmacol 141: 1077-1083. Either of theforegoing models can be adapted for use with other animals capable ofcoughing. Exemplary additional animals capable of coughing include catsand dogs.

Optimizing the Treatment of Pain

TRPV3 inhibitors, according to the present invention, can be used in thetreatment of a variety of injuries, diseases, conditions, and disorders.One important therapeutic use for TRPV3 inhibitors is in the treatmentof pain. As illustrated by the extensive list of injuries, conditions,and diseases for which pain is a significant and sometimes debilitatingsymptom, improved methods and compositions for use in the treatment ofpain provide substantial benefits for an enormous range of patients.Such methods and compositions have the potential to improve the qualityof care and the quality of life for patients afflicted with a diverserange of injuries, diseases, and conditions.

TRPV3 is a good target for modulating pain. TRPV3 is expressed intissues that contribute to transmission of painful stimuli.Additionally, TRPV3 expression is upregulated, for example in dorsalroot ganglia, following injury. Finally, TRPV3 knockout mice exhibitabnormal responses to painful stimuli. These characteristics of TRPV3suggest that inhibitors of TRPV3 will be useful in the treatment ofpain.

Many of these characteristics are shared by TRPV1, and inhibitors ofTRPV1 are being developed for the treatment of pain. However, althoughTRPV1 and TRPV3 share certain characteristics consistent with thedevelopment of effective therapeutics for the treatment of pain, TRPV3possesses certain characteristics that makes it a better target fortherapeutic compounds for the treatment of pain. For example, TRPV3sensitizes upon repeated stimulation. In contrast, TRPV1 desensitizesupon repeated stimulation with the agonist capsaicin. In addition toexpression in dorsal root ganglia, TRPV3 is expressed in skin. Given thesignificant involvement of skin in many types of pain, this expressionpattern is suggestive of potential effectiveness of TRPV3 inhibitors inpain involving the skin.

An important issue with the treatment of pain is how to manage painwhile reducing the side effects experienced with many analgesics. Forexample, although many opiates and other narcotics effectively diminishpain, patients are often unable to drive, work, or concentrate whiletaking these medications. Thus, while opiates such as morphine ordilaudin may be suitable for short term use or for use duringhospitalization, they are not optimal for long term use. Additionally,opiates and other narcotics are habit forming, and patients typicallydevelop a tolerance for these drugs. These characteristics of opioidsand other narcotics make them sub-optimal for pain management.

The present invention provides TRPV3 inhibitors for use in vitro and invivo. The present invention also provides compositions andpharmaceutical compositions comprising particular classes of compoundsthat inhibit TRPV3 activity. In certain embodiments, the subject TRPV3inhibitors are selective. In other words, in certain embodiments, thecompound inhibits TRPV3 activity preferentially over the activity ofother ion channels. In certain embodiments, the compound inhibits TRPV3activity preferentially over TRPV1, TRPV2, TRPV4, and/or TRPM8 activity.In certain other embodiments, the compound is selected because it crossreacts with one or more other TRP channels involved with pain. Forexample, in certain embodiments, the compound inhibits the activity ofTRPV3 and also inhibits the activity of one or more of TRPV1, TRPV2,TRPV4, and TRPM8.

Combination Therapy

Another aspect of the invention provides a conjoint therapy wherein oneor more other therapeutic agents are administered with the TRPV3modulators. Such conjoint treatment may be achieved by way of thesimultaneous, sequential, or separate dosing of the individualcomponents of the treatment.

In certain embodiments, a compound of the invention is conjointlyadministered with an analgesic. Suitable analgesics include, but are notlimited to, opioids, glucocorticosteroids, non-steroidalanti-inflammatories, naphthylalkanones, oxicams, para-aminophenolderivatives, propionic acids, propionic acid derivatives, salicylates,fenamates, fenamate derivatives, pyrozoles, and pyrozole derivatives.Examples of such analgesic compounds include, but are not limited to,codeine, hydrocodone, hydromorphone, levorpharnol, morphine, oxycodone,oxymorphone, butorphanol, dezocine, nalbuphine, pentazocine, etodolac,indomethacin, sulindac, tolmetin, nabumetone, piroxicam, acetaminophen,fenoprofen, flurbiprofen, ibuprofen, ketoprofen, naproxen, diclofenac,oxaprozin, aspirin, diflunisal, meclofenamic acid, mefanamic acid,prednisolone, and dexamethasone. Preferred analgesics are non-steroidalanti-inflammatories and opioids (preferably morphine).

In certain embodiments, a compound of the invention is conjointlyadministered with a non-steroidal anti-inflammatory. Suitablenon-steroidal anti-inflammatory compounds include, but are not limitedto, piroxicam, diclofenac, etodolac, indomethacin, ketoralac, oxaprozin,tolmetin, naproxen, flubiprofen, fenoprofen, ketoprofen, ibuprofen,mefenamic acid, sulindac, apazone, phenylbutazone, aspirin, celecoxiband rofecoxib.

In certain embodiments, a compound of the invention is conjointlyadministered with an antiviral agent. Suitable antiviral agents include,but are not limited to, amantadine, acyclovir, cidofovir, desciclovir,deoxyacyclovir, famciclovir, foscamet, ganciclovir, penciclovir,azidouridine, anasmycin, amantadine, bromovinyldeoxusidine,chlorovinyldeoxusidine, cytarbine, didanosine, deoxynojirimycin,dideoxycitidine, dideoxyinosine, dideoxynucleoside, edoxuidine,enviroxime, fiacitabine, foscamet, fialuridine, fluorothymidine,floxuridine, hypericin, interferon, interleukin, isethionate,nevirapine, pentarnidine, ribavirin, rimantadine, stavirdine,sargramostin, suramin, trichosanthin, tribromothymidine,trichlorothymidine, vidarabine, zidoviridine, zalcitabine3-azido-3-deoxythymidine, 2′,3′-dideoxyadenosine (ddA),2′,3′-dideoxyguanosine (ddG), 2′,3′-dideoxycytidine (ddC),2′,3′-dideoxythymidine (ddT), 2′3′-dideoxy-dideoxythymidine (d4T),2′-deoxy-3′-thia-cytosine (3TC or lamivudime),2′,3′-dideoxy-2′-fluoroadenosine, 2′,3′-dideoxy-2′-fluoroinosine,2′,3′-dideoxy-2′-fluorothymidine, 2′,3′-dideoxy-2′-fluorocytosine,2′3′-dideoxy-2′,3′-didehydro-2′-fluorothymidine (Fd4T),2′3′-dideoxy-2′-beta-fluoroadenosine (F-ddA),2′3′-dideoxy-2′-beta-fluoro-inosine (F-ddI), and2′,3′-dideoxy-2′-beta-flurocytosine (F-ddC), trisodiumphosphomonoformate, trifluorothymidine, 3′azido-3′thymidine (AZT),dideoxyinosine (ddI), and idoxuridine.

In certain embodiments, a compound of the invention is conjointlyadministered with an antibacterial agent. Suitable antibacterial agentsinclude, but are not limited to, amanfadine hydrochloride, amanfadinesulfate, amikacin, amikacin sulfate, amoglycosides, amoxicillin,ampicillin, amsamycins, bacitracin, beta-lactams, candicidin,capreomycin, carbenicillin, cephalexin, cephaloridine, cephalothin,cefazolin, cephapirin, cephradine, cephaloglycin, chilomphenicols,chlorhexidine, chloshexidine gluconate, chlorhexidine hydrochloride,chloroxine, chlorquiraldol, chlortetracycline, chlortetracyclinehydrochloride, ciprofloxacin, circulin, clindamycin, clindamycinhydrochloride, clotrimazole, cloxacillin, demeclocycline,diclosxacillin, diiodohydroxyquin, doxycycline, ethambutol, ethambutolhydrochloride, erythromycin, erythromycin estolate, erhmycin stearate,farnesol, floxacillin, gentamicin, gentamicin sulfate, gramicidin,giseofulvin, haloprogin, haloquinol, hexachlorophene, iminocylcline,iodochlorhydroxyquin, kanamycin, kanamycin sulfate, lincomycin,lineomycin, lineomycin hydrochoride, macrolides, meclocycline,methacycline, methacycline hydrochloride, methenine, methenaminehippurate, methenamine mandelate, methicillin, metonidazole, miconazole,miconazole hydrochloride, minocycline, minocycline hydrochloride,mupirocin, nafcillin, neomycin, neomycin sulfate, netimicin, netimicinsulfate, nitrofurazone, norfloxacin, nystatin, octopirox, oleandomycin,orcephalosporins, oxacillin, oxyteacline, oxytetracycline hydrochloride,parachlorometa xylenol, paromomycin, paromomycin sulfate, penicillins,penicillin G, penicillin V, pentamidine, pentamidine hydrochloride,phenethicillin, polymyxins, quinolones, streptomycin sulfate,tetracycline, tobramycin, tolnaftate, triclosan, trifampin, rifamycin,rolitetracycline, spectinomycin, spiramycin, struptomycin, sulfonamide,tetracyclines, tetracycline, tobramycin, tobramycin sulfate,triclocarbon, triclosan, trimethoprim-sulfamethoxazole, tylosin,vancomycin, and yrothricin.

In certain embodiments, a compound of the invention is conjointlyadministered with a cough suppressant, decongestant, or expectorant.

Examples of retinoids that be administered with the subject TRPV3inhibitors, e.g., where the TRPV3 inhibitor can be used to reduce thepain and/or inflammatory effect of the retinoid, include, but are notlimited to, compounds such as retinoic acid (both cis and trans),retinol, adapalene, vitamin A and tazarotene. Retinoids are useful intreating acne, psoriasis, rosacea, wrinkles and skin cancers and cancerprecursors such as melanoma and actinic keratosis.

Similarly, the subject TRPV3 inhibitors can be used in conjunction withkeratolytic agents include benzoyl peroxide, alpha hydroxyacids, fruitacids, glycolic acid, salicylic acid, azelaic acid, trichloroaceticacid, lactic acid and piroctone.

The subject TRPV3 inhibitors can also be administered along withdepilatory agents (hair loss).

The subject TRPV3 inhibitors can be used with anti-acne agents,anti-eczema agents and anti-psoratic agents. Compounds particlarlyuseful in treating acne include azelaic acid (an aliphatic diacid withantiacne properties), anthralin (a diphenolic compound with antifungaland antipsoriatic properties), and masoprocol (nordihydroguaiareticacid, a tetraphenolic compound with antioxidant properties, also usefulin the treatment of actinic keratosis) and analogs thereof (such asaustrobailignan 6, oxoaustrobailignan6,4′-O-methyl-7,7′-dioxoaustrobailignan 6, macelignan,demethyldihydroguaiaretic acid, 3,3′,4-trihydroxy-4′-methoxylignan,Saururenin, 4-hydroxy-3,3′,4′-trimethoxylignan, and isoanwulignan).Anti-eczema agents include pimecrolimus and tacrolimus. Anti-psoriaticactive agents suitable for use in the present invention includeretinoids (including isomers and derivatives of retinoic acid, as wellas other compounds that bind to the retinoic acid receptor, such asretinoic acid, acitretin, 13-cis-retinoic acid (isotretinoin),9-cis-retinoic acid, tocopheryl-retinoate (tocopherol ester of retinoicacid (trans- or cis-)), etretinate, motretinide,1-(13-cis-retinoyloxy)-2-propanone,1-(13-cis-retinoyloxy)-3-decanoyloxy-2-propanone,1,3-bis-(13-cis-retinoyloxy)-2-propanone,2-(13-cis-retinoyloxy)-acetophenone,13-cis-retinoyloxymethyl-2,2-dimethyl propanoate,2-(13-cis-retinoyloxy)-n-methyl-acetamide,1-(13-cis-retinoyloxy)-3-hydroxy-2-propanone,1-(13-cis-retinoyloxy)-2,3-dioleoylpropanone, succinimdyl13-cis-retinoate, adapalene, and tazarotene), salicylic acid(monoammonium salt), anthralin, 6-azauridine, vitamin D derivatives(including but not limited to Rocaltrol (Roche Laboratories), EB 1089(24α,26α,27α-trihomo-22,24-diene-1α,25-(OH)₂-D₃), KH 1060(20-epi-22-oxa-24α,26α,27α-trihomo-1α,25-(OH)₂-D₃), MC 1288, GS 1558, CB1093, 1,25-(OH)₂-16-ene-D₃, 1,25-(OH)₂-16-ene-23-yne-D₃, and25-(OH)2-16-ene-23-yne-D₃, 22-oxacalcitriol; 1α-(OH)D₅ (University ofIllinois), ZK 161422 and ZK 157202 (Institute of MedicalChemistry-Schering AG), alfacalcidol, calcifediol, calcipotriol(calcipotriene), maxacalcitriol, colecalciferol, doxercalciferol,ergocalciferol, falecalcitriol, lexacalcitol, maxacalcitol,paricalcitol, secalciferol, seocalcitol, tacalcitol, calcipotriene,calcitriol, and other analogs as disclosed in U.S. Pat. No. 5,994,332),pyrogallol, and tacalcitol.

The subject TRPV3 inhibitors can also be administered with vitamins andderivatives thereof including Vitamin A, ascorbic acid (Vitamin C),alpha-tocopherol (Vitamin E), 7-dehydrocholesterol (Vitamin D), VitaminK, alpha-lipoic acid, lipid soluble anti-oxidants, and the like.

The subject TRPV3 inhibitors can also be used with skin protectants,such allantoin and esculin.

In certain embodiments, two or more compounds of the invention areconjointly administered. When two or more compounds of the invention areconjointly administered, the two or more compounds may have a similarselectivity profile and functional activity, or the two or morecompounds may have a different selectivity profile and functionalactivity. By way of example, the two or more compounds may both beapproximately 10, 100, or 1000 fold selective for antagonizing afunction of TRPV3 over TRPV1, TRPV5, and TRPV6 (e.g., the two or morecompounds have a similar selectivity profile), and further may inhibit afunction of TRPV3 with a similar IC₅₀ (e.g., a similar functionalactivity). Alternatively, the one of the two or more compounds mayselectively inhibit TRPV3 while the other of the two or more compoundsinhibits both TRPV3 and TRPV1 (e.g., the two or more compounds havediffering selectivity profiles). Administration of combinations of twoor more compounds of the invention having similar or differingproperties are contemplated.

In certain embodiments, a compound of the invention is conjointlyadministered with one or more additional compounds that antagonize thefunction of a different channel. By way of example, a compound of theinvention may be conjointly administered with one or more compounds thatantagonize TRPV1 and/or TRPV4. The compound(s) that antagonize TRPV1 orTRPV4 may be selective for TRPV1 or TRPV4 (e.g, inhibit TRPV1 or TRPV410, 100, or 1000 fold more strongly than TRPV3). Alternatively, thecompound(s) that antagonize TRPV1 or TRPV4 may cross react with otherTRP channels.

In certain other embodiments, a compound of the invention is conjointlyadministered with one or more additional agents or therapeutic regimensappropriate for the particular injury, disease, condition, or disorderbeing treated.

When combinations of a TRPV3 inhibitor and one or more other compounds,agents, or therapeutic regimens are administered, the inventioncontemplates administration via the same route of administration or viadiffering routes of administration.

Pharmaceutical Compositions

While it is possible for a compound of the present invention to beadministered alone, it is preferable to administer the compound as apharmaceutical formulation (composition). The compounds according to theinvention may be formulated for administration in any convenient way foruse in human or veterinary medicine. In certain embodiments, thecompound included in the pharmaceutical preparation may be activeitself, or may be a prodrug, e.g., capable of being converted to anactive compound in a physiological setting.

Regardless of the route of administration selected, the compounds of thepresent invention, which may be used in a suitable hydrated form, and/orthe pharmaceutical compositions of the present invention, are formulatedinto pharmaceutically acceptable dosage forms such as described below orby other conventional methods known to those of skill in the art.

Thus, another aspect of the present invention provides pharmaceuticallyacceptable compositions comprising a therapeutically effective amount ofone or more of the compounds described above, formulated together withone or more pharmaceutically acceptable carriers (additives) and/ordiluents. As described in detail below, the pharmaceutical compositionsof the present invention may be specially formulated for administrationin solid or liquid form, including those adapted for the following: (1)oral administration, for example, drenches (aqueous or non-aqueoussolutions or suspensions); tablets; boluses; powders; granules; pastesfor application to the tongue, teeth, lips, gums; mouth washes; gels;(2) parenteral administration, for example, by subcutaneous,intramuscular or intravenous injection as, for example, a sterilesolution or suspension; (3) topical application, for example, as acream, ointment or spray applied to the skin; (4) intravaginally orintrarectally, for example, as a pessary, cream or foam; or (5) forinhalation. However, in certain embodiments the subject compounds may besimply dissolved or suspended in sterile water. In certain embodiments,the pharmaceutical preparation is non-pyrogenic, i.e., does not elevatethe body temperature of a patient.

The phrase “therapeutically effective amount” as used herein means thatamount of a compound, material, or composition comprising a compound ofthe present invention which is effective for producing some desiredtherapeutic effect by inhibiting TRPV3 function in at least asub-population of cells in an animal and thereby blocking the biologicalconsequences of that function in the treated cells, at a reasonablebenefit/risk ratio applicable to any medical treatment.

The phrases “systemic administration,” “administered systemically,”“peripheral administration” and “administered peripherally” as usedherein mean the administration of a compound, drug or other materialother than directly into the central nervous system, such that it entersthe patient's system and, thus, is subject to metabolism and other likeprocesses, for example, subcutaneous administration.

The phrase “pharmaceutically acceptable” is employed herein to refer tothose compounds, materials, compositions, and/or dosage forms which are,within the scope of sound medical judgment, suitable for use in contactwith the tissues of human beings and animals without excessive toxicity,irritation, allergic response, or other problem or complication,commensurate with a reasonable benefit/risk ratio.

The phrase “pharmaceutically acceptable carrier” as used herein means apharmaceutically acceptable material, composition or vehicle, such as aliquid or solid filler, diluent, excipient, solvent or encapsulatingmaterial, involved in carrying or transporting the subject antagonistsfrom one organ, or portion of the body, to another organ, or portion ofthe body. Each carrier must be “acceptable” in the sense of beingcompatible with the other ingredients of the formulation and notinjurious to the patient. Some examples of materials which can serve aspharmaceutically acceptable carriers include: (1) sugars, such aslactose, glucose and sucrose; (2) starches, such as corn starch andpotato starch; (3) cellulose, and its derivatives, such as sodiumcarboxymethyl cellulose, ethyl cellulose and cellulose acetate; (4)powdered tragacanth; (5) malt; (6) gelatin; (7) talc; (8) excipients,such as cocoa butter and suppository waxes; (9) oils, such as peanutoil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil andsoybean oil; (10) glycols, such as propylene glycol; (11) polyols, suchas glycerin, sorbitol, mannitol and polyethylene glycol; (12) esters,such as ethyl oleate and ethyl laurate; (13) agar; (14) bufferingagents, such as magnesium hydroxide and aluminum hydroxide; (15) alginicacid; (16) pyrogen-free water; (17) isotonic saline; (18) Ringer'ssolution; (19) ethyl alcohol; (20) phosphate buffer solutions; and (21)other non-toxic compatible substances employed in pharmaceuticalformulations.

As set out above, certain embodiments of the present compounds maycontain a basic functional group, such as amino or alkylamino, and are,thus, capable of forming pharmaceutically acceptable salts withpharmaceutically acceptable acids. The term “pharmaceutically acceptablesalts” in this respect, refers to the relatively non-toxic, inorganicand organic acid addition salts of compounds of the present invention.These salts can be prepared in situ during the final isolation andpurification of the compounds of the invention, or by separatelyreacting a purified compound of the invention in its free base form witha suitable organic or inorganic acid, and isolating the salt thusformed. Representative salts include the hydrobromide, hydrochloride,sulfate, bisulfate, phosphate, nitrate, acetate, valerate, oleate,palmitate, stearate, laurate, benzoate, lactate, phosphate, tosylate,citrate, maleate, fumarate, succinate, tartrate, napthylate, mesylate,glucoheptonate, lactobionate, and laurylsulphonate salts and the like.(See, for example, Berge et al. (1977) “Pharmaceutical Salts”, J. Pharm.Sci. 66:1-19)

The pharmaceutically acceptable salts of the subject compounds includethe conventional nontoxic salts or quaternary ammonium salts of thecompounds, e.g., from non-toxic organic or inorganic acids. For example,such conventional nontoxic salts include those derived from inorganicacids such as hydrochloride, hydrobromic, sulfuric, sulfamic,phosphoric, nitric, and the like; and the salts prepared from organicacids such as acetic, propionic, succinic, glycolic, stearic, lactic,malic, tartaric, citric, ascorbic, palmitic, maleic, hydroxymaleic,phenylacetic, glutamic, benzoic, salicyclic, sulfanilic,2-acetoxybenzoic, fumaric, toluenesulfonic, methanesulfonic, ethanedisulfonic, oxalic, isothionic, and the like.

In other cases, the compounds of the present invention may contain oneor more acidic functional groups and, thus, are capable of formingpharmaceutically acceptable salts with pharmaceutically acceptablebases. The term “pharmaceutically acceptable salts” in these instancesrefers to the relatively non-toxic, inorganic and organic base additionsalts of compounds of the present invention. These salts can likewise beprepared in situ during the final isolation and purification of thecompounds, or by separately reacting the purified compound in its freeacid form with a suitable base, such as the hydroxide, carbonate orbicarbonate of a pharmaceutically acceptable metal cation, with ammonia,or with a pharmaceutically acceptable organic primary, secondary ortertiary amine. Representative alkali or alkaline earth salts includethe lithium, sodium, potassium, calcium, magnesium, and aluminum saltsand the like. Representative organic amines useful for the formation ofbase addition salts include ethylamine, diethylamine, ethylenediamine,ethanolamine, diethanolamine, piperazine and the like. (See, forexample, Berge et al., supra)

Wetting agents, emulsifiers and lubricants, such as sodium laurylsulfate and magnesium stearate, as well as coloring agents, releaseagents, coating agents, sweetening, flavoring and perfuming agents,preservatives and antioxidants can also be present in the compositions.

Examples of pharmaceutically acceptable antioxidants include: (1) watersoluble antioxidants, such as ascorbic acid, cysteine hydrochloride,sodium bisulfate, sodium metabisulfite, sodium sulfite and the like; (2)oil-soluble antioxidants, such as ascorbyl palmitate, butylatedhydroxyanisole (BHA), butylated hydroxytoluene (BHT), lecithin, propylgallate, alpha-tocopherol, and the like; and (3) metal chelating agents,such as citric acid, ethylenediamine tetraacetic acid (EDTA), sorbitol,tartaric acid, phosphoric acid, and the like.

Formulations of the present invention include those suitable for oral,nasal, topical (including buccal and sublingual), rectal, vaginal and/orparenteral administration. Oral formulations include those delivered toand maintained in the mouth without swallowing, as well as formulationsthat are swallowed as part of or following use. The formulations mayconveniently be presented in unit dosage form and may be prepared by anymethods well known in the art of pharmacy. The amount of activeingredient which can be combined with a carrier material to produce asingle dosage form will vary depending upon the host being treated, theparticular mode of administration. The amount of active ingredient thatcan be combined with a carrier material to produce a single dosage formwill generally be that amount of the compound which produces atherapeutic effect. Generally, out of one hundred percent, this amountwill range from about 1 percent to about ninety-nine percent of activeingredient, preferably from about 5 percent to about 70 percent, mostpreferably from about 10 percent to about 30 percent.

Methods of preparing these formulations or compositions include the stepof bringing into association a compound of the present invention withthe carrier and, optionally, one or more accessory ingredients. Ingeneral, the formulations are prepared by uniformly and intimatelybringing into association a compound of the present invention withliquid carriers, or finely divided solid carriers, or both, and then, ifnecessary, shaping the product.

Formulations of the invention suitable for oral administration may be inthe form of capsules, cachets, pills, tablets, lozenges (using aflavored basis, usually sucrose and acacia or tragacanth), powders,granules, or as a solution or a suspension in an aqueous or non-aqueousliquid, or as an oil-in-water or water-in-oil liquid emulsion, or as anelixir or syrup, or as pastilles (using an inert base, such as gelatinand glycerin, or sucrose and acacia) and/or as mouth washes and thelike, each containing a predetermined amount of a compound of thepresent invention as an active ingredient. A compound of the presentinvention may also be administered as a bolus, electuary or paste.

In solid dosage forms of the invention for oral administration(capsules, tablets, pills, dragees, powders, granules and the like), theactive ingredient is mixed with one or more pharmaceutically acceptablecarriers, such as sodium citrate or dicalcium phosphate, and/or any ofthe following: (1) fillers or extenders, such as starches, lactose,sucrose, glucose, mannitol, and/or silicic acid; (2) binders, such as,for example, carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidone, sucrose and/or acacia; (3) humectants, such as glycerol;(4) disintegrating agents, such as agar-agar, calcium carbonate, potatoor tapioca starch, alginic acid, certain silicates, and sodiumcarbonate; (5) solution retarding agents, such as paraffin; (6)absorption accelerators, such as quaternary ammonium compounds; (7)wetting agents, such as, for example, cetyl alcohol and glycerolmonostearate; (8) absorbents, such as kaolin and bentonite clay; (9)lubricants, such a talc, calcium stearate, magnesium stearate, solidpolyethylene glycols, sodium lauryl sulfate, and mixtures thereof; and(10) coloring agents. In the case of capsules, tablets and pills, thepharmaceutical compositions may also comprise buffering agents. Solidcompositions of a similar type may also be employed as fillers in softand hard-filled gelatin capsules using such excipients as lactose ormilk sugars, as well as high molecular weight polyethylene glycols andthe like.

A tablet may be made by compression or molding, optionally with one ormore accessory ingredients. Compressed tablets may be prepared usingbinder (for example, gelatin or hydroxypropylmethyl cellulose),lubricant, inert diluent, preservative, disintegrant (for example,sodium starch glycolate or cross-linked sodium carboxymethyl cellulose),surface-active or dispersing agent. Molded tablets may be made bymolding in a suitable machine a mixture of the powdered compoundmoistened with an inert liquid diluent.

The tablets, and other solid dosage forms of the pharmaceuticalcompositions of the present invention, such as dragees, capsules, pillsand granules, may optionally be scored or prepared with coatings andshells, such as enteric coatings and other coatings well known in thepharmaceutical-formulating art. They may also be formulated so as toprovide slow or controlled release of the active ingredient thereinusing, for example, hydroxypropylmethyl cellulose in varying proportionsto provide the desired release profile, other polymer matrices,liposomes and/or microspheres. They may be sterilized by, for example,filtration through a bacteria-retaining filter, or by incorporatingsterilizing agents in the form of sterile solid compositions that can bedissolved in sterile water, or some other sterile injectable mediumimmediately before use. These compositions may also optionally containopacifying agents and may be of a composition that they release theactive ingredient(s) only, or preferentially, in a certain portion ofthe gastrointestinal tract, optionally, in a delayed manner. Examples ofembedding compositions that can be used include polymeric substances andwaxes. The active ingredient can also be in micro-encapsulated form, ifappropriate, with one or more of the above-described excipients.

Liquid dosage forms for oral administration of the compounds of theinvention include pharmaceutically acceptable emulsions, microemulsions,solutions, suspensions, syrups and elixirs. In addition to the activeingredient, the liquid dosage forms may contain inert diluents commonlyused in the art, such as, for example, water or other solvents,solubilizing agents and emulsifiers, such as ethyl alcohol, isopropylalcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzylbenzoate, propylene glycol, 1,3-butylene glycol, oils (in particular,cottonseed, groundnut, corn, germ, olive, castor and sesame oils),glycerol, tetrahydrofuryl alcohol, polyethylene glycols and fatty acidesters of sorbitan, and mixtures thereof.

Besides inert diluents, the oral compositions can also include adjuvantssuch as wetting agents, emulsifying and suspending agents, sweetening,flavoring, coloring, perfuming and preservative agents.

Suspensions, in addition to the active compounds, may contain suspendingagents as, for example, ethoxylated isostearyl alcohols, polyoxyethylenesorbitol and sorbitan esters, microcrystalline cellulose, aluminummetahydroxide, bentonite, agar-agar and tragacanth, and mixturesthereof.

Formulations for oral administration may be administered directly to themouth in the presence or absence of a device to aid administration orlocal delivery. For example, a liquid formulation or suspension may bedirectly delivered via a mouthwash. Alternatively, the liquidformulation or suspension may be directly applied to all or a portion ofthe mouth using a syringe or swab. In another embodiment, an oralformulation may be applied to a mouth guard or other dental device, anddelivered to the mouth via the mouth guard or device. The presentinvention contemplates that preparations suitable for oral delivery canbe formulated to facilitate any of these modes of delivery. For any ofthe foregoing, the oral formulation may optionally be ingested or may bemaintained in the mouth and later expectorated.

It is known that sterols, such as cholesterol, will form complexes withcyclodextrins. Thus, in preferred embodiments, where the inhibitor is asteroidal alkaloid, it may be formulated with cyclodextrins, such as α-,β- and γ-cyclodextrin, dimethyl-β cyclodextrin and2-hydroxypropyl-β-cyclodextrin.

Formulations of the pharmaceutical compositions of the invention forrectal or vaginal administration may be presented as a suppository,which may be prepared by mixing one or more compounds of the inventionwith one or more suitable nonirritating excipients or carrierscomprising, for example, cocoa butter, polyethylene glycol, asuppository wax or a salicylate, and which is solid at room temperature,but liquid at body temperature and, therefore, will melt in the rectumor vaginal cavity and release the active compound.

Formulations of the present invention which are suitable for vaginaladministration also include pessaries, tampons, creams, gels, pastes,foams or spray formulations containing such carriers as are known in theart to be appropriate.

Dosage forms for the topical or transdermal administration of a compoundof this invention include powders, sprays, ointments, pastes, creams,lotions, gels, solutions, patches and inhalants. The active compound maybe mixed under sterile conditions with a pharmaceutically acceptablecarrier, and with any preservatives, buffers, or propellants that may berequired.

The ointments, pastes, creams and gels may contain, in addition to anactive compound of this invention, excipients, such as animal andvegetable fats, oils, waxes, paraffins, starch, tragacanth, cellulosederivatives, polyethylene glycols, silicones, bentonites, silicic acid,talc and zinc oxide, or mixtures thereof.

Powders and sprays can contain, in addition to a compound of thisinvention, excipients such as lactose, talc, silicic acid, aluminumhydroxide, calcium silicates and polyamide powder, or mixtures of thesesubstances. Sprays can additionally contain customary propellants, suchas chlorofluorohydrocarbons and volatile unsubstituted hydrocarbons,such as butane and propane.

Transdermal patches have the added advantage of providing controlleddelivery of a compound of the present invention to the body. Such dosageforms can be made by dissolving or dispersing the compound in the propermedium. Absorption enhancers can also be used to increase the flux ofthe compound across the skin. The rate of such flux can be controlled byeither providing a rate controlling membrane or dispersing the compoundin a polymer matrix or gel.

Ophthalmic formulations, eye ointments, powders, solutions and the like,are also contemplated as being within the scope of this invention.

The phrases “parenteral administration” and “administered parenterally”as used herein means modes of administration other than enteral andtopical administration, usually by injection, and includes, withoutlimitation, intravenous, intramuscular, intraarterial, intrathecal,intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal,transtracheal, subcutaneous, subcuticular, intraarticular, subcapsular,subarachnoid, intraspinal and intrasternal injection and infusion.

Pharmaceutical compositions of this invention suitable for parenteraladministration comprise one or more compounds of the invention incombination with one or more pharmaceutically acceptable sterileisotonic aqueous or nonaqueous solutions, dispersions, suspensions oremulsions, or sterile powders which may be reconstituted into sterileinjectable solutions or dispersions just prior to use, which may containantioxidants, buffers, bacteriostats, solutes which render theformulation isotonic with the blood of the intended recipient orsuspending or thickening agents.

Examples of suitable aqueous and nonaqueous carriers that may beemployed in the pharmaceutical compositions of the invention includewater, ethanol, polyols (such as glycerol, propylene glycol,polyethylene glycol, and the like), and suitable mixtures thereof,vegetable oils, such as olive oil, and injectable organic esters, suchas ethyl oleate. Proper fluidity can be maintained, for example, by theuse of coating materials, such as lecithin, by the maintenance of therequired particle size in the case of dispersions, and by the use ofsurfactants.

These compositions may also contain adjuvants such as preservatives,wetting agents, emulsifying agents and dispersing agents. Prevention ofthe action of microorganisms may be ensured by the inclusion of variousantibacterial and antifungal agents, for example, paraben,chlorobutanol, phenol sorbic acid, and the like. It may also bedesirable to include isotonic agents, such as sugars, sodium chloride,and the like into the compositions. In addition, prolonged absorption ofthe injectable pharmaceutical form may be brought about by the inclusionof agents that delay absorption such as aluminum monostearate andgelatin.

In some cases, in order to prolong the effect of a drug, it is desirableto slow the absorption of the drug from subcutaneous or intramuscularinjection. This may be accomplished by the use of a liquid suspension ofcrystalline or amorphous material having poor water solubility. The rateof absorption of the drug then depends upon its rate of dissolution,which, in turn, may depend upon crystal size and crystalline form.Alternatively, delayed absorption of a parenterally administered drugform is accomplished by dissolving or suspending the drug in an oilvehicle.

Injectable depot forms are made by forming microencapsule matrices ofthe subject compounds in biodegradable polymers such aspolylactide-polyglycolide. Depending on the ratio of drug to polymer,and the nature of the particular polymer employed, the rate of drugrelease can be controlled. Examples of other biodegradable polymersinclude poly(orthoesters) and poly(anhydrides). Depot injectableformulations are also prepared by entrapping the drug in liposomes ormicroemulsions that are compatible with body tissue.

When the compounds of the present invention are administered aspharmaceuticals, to humans and animals, they can be given per se or as apharmaceutical composition containing, for example, 0.1 to 99.5% (morepreferably, 0.5 to 90%) of active ingredient in combination with apharmaceutically acceptable carrier.

The addition of the active compound of the invention to animal feed ispreferably accomplished by preparing an appropriate feed premixcontaining the active compound in an effective amount and incorporatingthe premix into the complete ration.

Alternatively, an intermediate concentrate or feed supplement containingthe active ingredient can be blended into the feed. The way in whichsuch feed premixes and complete rations can be prepared and administeredare described in reference books (such as “Applied Animal Nutrition”,W.H. Freedman and CO., San Francisco, U.S.A., 1969 or “Livestock Feedsand Feeding” O and B books, Corvallis, Oreg., U.S.A., 1977).

Methods of introduction may also be provided by rechargeable orbiodegradable devices. Various slow release polymeric devices have beendeveloped and tested in vivo in recent years for the controlled deliveryof drugs, including proteinacious biopharmaceuticals. A variety ofbiocompatible polymers (including hydrogels), including bothbiodegradable and non-degradable polymers, can be used to form animplant for the sustained release of a compound at a particular targetsite.

Actual dosage levels of the active ingredients in the pharmaceuticalcompositions of this invention may be varied so as to obtain an amountof the active ingredient that is effective to achieve the desiredtherapeutic response for a particular patient, composition, and mode ofadministration, without being toxic to the patient.

The selected dosage level will depend upon a variety of factorsincluding the activity of the particular compound of the presentinvention employed, or the ester, salt or amide thereof, the route ofadministration, the time of administration, the rate of excretion of theparticular compound being employed, the duration of the treatment, otherdrugs, compounds and/or materials used in combination with theparticular compound employed, the age, sex, weight, condition, generalhealth and prior medical history of the patient being treated, and likefactors well known in the medical arts.

A physician or veterinarian having ordinary skill in the art can readilydetermine and prescribe the effective amount of the pharmaceuticalcomposition required. For example, the physician or veterinarian couldstart doses of the compounds of the invention employed in thepharmaceutical composition at levels lower than that required in orderto achieve the desired therapeutic effect and gradually increase thedosage until the desired effect is achieved.

In general, a suitable daily dose of a compound of the invention will bethat amount of the compound that is the lowest dose effective to producea therapeutic effect. Such an effective dose will generally depend uponthe factors described above. Generally, intravenous,intracerebroventricular and subcutaneous doses of the compounds of thisinvention for a patient will range from about 0.0001 to about 100 mg perkilogram of body weight per day.

If desired, the effective daily dose of the active compound may beadministered as two, three, four, five, six or more sub-dosesadministered separately at appropriate intervals throughout the day,optionally, in unit dosage forms.

The patient receiving this treatment is any animal in need, includingprimates, in particular humans, and other mammals such as equines,cattle, swine and sheep; and poultry and pets in general.

The compound of the invention can be administered as such or inadmixtures with pharmaceutically acceptable and/or sterile carriers andcan also be administered in conjunction with other agents. Exemplaryother agents include penicillins, cephalosporins, aminoglycosides,glycopeptides, anti-inflammatories, anti-virals, anti-fungals,anti-bacterials, or any agents appropriate for the treatment of theparticular injury, disease, or condition. Conjunctive therapy, thusincludes sequential, simultaneous and separate administration of theactive compound in a way that the therapeutic effects of the firstadministered compound are still detectable when the subsequent therapyis administered.

Synthetic Schemes and Identification of Active Antagonists

Combinatorial Libraries

The compounds of the present invention, particularly libraries ofvariants having various representative classes of substituents, areamenable to combinatorial chemistry and other parallel synthesis schemes(see, for example, PCT WO 94/08051). The result is that large librariesof related compounds, e.g. a variegated library of compounds representedabove, can be screened rapidly in high throughput assays in order toidentify potential TRPV3 agonist or antagonist lead compounds, as wellas to refine the specificity, toxicity, and/or cytotoxic-kinetic profileof a lead compound. For instance, TRPV3 bioactivity assays, such asthose disclosed herein, can be used to screen a library of compounds forthose having agonist activity or antagonist activity towards TRPV3.

Simply for illustration, a combinatorial library for the purposes of thepresent invention is a mixture of chemically related compounds that maybe screened together for a desired property. The preparation of manyrelated compounds in a single reaction greatly reduces and simplifiesthe number of screening processes that need to be carried out. Screeningfor the appropriate physical properties can be done by conventionalmethods.

Diversity in the library can be created at a variety of differentlevels. For instance, the substrate aryl groups used in thecombinatorial reactions can be diverse in terms of the core aryl moiety,e.g., a variegation in terms of the ring structure, and/or can be variedwith respect to the other substituents.

A variety of techniques are available in the art for generatingcombinatorial libraries of small organic molecules such as the subjectcompounds. See, for example, Blondelle et al. (1995) Trends Anal. Chem.14:83; the Affymax U.S. Pat. Nos. 5,359,115 and 5,362,899: the EllmanU.S. Pat. No. 5,288,514: the Still et al. PCT publication WO 94/08051;the ArQule U.S. Pat. Nos. 5,736,412 and 5,712,171; Chen et al. (1994)JACS 116:2661: Kerr et al. (1993) JACS 115:252; PCT publicationsWO92/10092, WO93/09668 and WO91/07087; and the Lerner et al. PCTpublication WO93/20242). Accordingly, a variety of libraries on theorder of about 100 to 1,000,000 or more diversomers of the subjectcompounds can be synthesized and screened for particular activity orproperty.

Many variations on the above and related pathways permit the synthesisof widely diverse libraries of compounds that may be tested asinhibitors or agonists of TRPV3.

EXAMPLES Example 1 Synthesis of Compounds of the Invention Synthesis of2-Chloro-1-(8-methyl-3,4-dihydro-2H-quinolin-1-yl)ethanone (2)

2-Chloro-1-(8-methyl-3,4-dihydro-2H-quinolin-1-yl)ethanone (2): To asolution of 8-methyl-1,2,3,4-tetrahydroquinoline (1) (2.77 g, 18.8 mmol)in dry CH₂Cl₂ (50 mL) at 0° C. was added chloroacetyl chloride (1.80 mL,22.6 mmol) followed by dropwise addition of Et₃N (6.30 mL, 45.2 mmol).The reaction was stirred overnight under N₂. The reaction mixture wasthen diluted with CH₂Cl₂ (50 mL), washed with water (2×15 mL), andsaturated NaCl (15 mL). Dried over MgSO₄, filtered, and concentrated togive compound 2 (3.7 g). This crude material was used in subsequentreactions without further purification.

Synthesis of 2-Chloro-1-(3,4-dihydro-2H-quinolin-1-yl)ethanone (4)

2-Chloro-1-(3,4-dihydro-2H-quinolin-1-yl)ethanone (4): To a solution of1,2,3,4-tetrahydroquinoline (3, 2.00 g, 15.0 mmol) in dry CH₂Cl₂ (30 mL)at 0° C. was added chloroacetyl chloride (1.80 g, 16.0 mmol) followed bydropwise addition of Et₃N (2.50 mL, 18.0 mmol). The reaction was stirredovernight under N₂. The reaction mixture was diluted with CH₂Cl₂ (50mL), washed with 5% HCl (15 mL), water (2×15 mL), and saturated NaCl (15mL). Dried over MgSO₄, filtered, and concentrated to give 3.05 g of 4.This crude material was used in subsequent reactions without furtherpurification.

Synthesis of 2,3,4,5-Tetrahydro-1H-benzo[b]azepine

2,3,4,5-Tetrahydro-1H-benzo[b]azepine (6): A mixture of1,2,3,4-tetrahydro-benzo[b]azepin-5-one (5) (1.58 g, 9.80 mmol),hydrazine monohydrate (1.92 mL, 40.0 mmol), and KOH (3.3 g, 58.8 mmol)in diethylene glycol (50 mL) was heated to reflux for 6 hr under a N₂atmosphere. After cooling, the reaction was poured onto ice-cold water,neutralized with 1N HCl, and extracted with EtOAc. All the organics werewashed with water and saturated NaCl. Dried over MgSO₄, filtered, andconcentrated to yield compound 6 (0.69 g). This crude material was usedin the next step with out further purification.

Synthesis of 5,6-Dichloro-2-mercapto benzothiazole

5,6-Dichloro-2-mercapto benzothiazole (8): To a suspension of CS₂ (0.64mL, 10.6 mmol) and sodium methylate (0.55 g, 10.2 mmol) in DMF (10 mL)was added 2,4,5-trichloroaniline (7, 1.00 g, 5.09 mmol) in DMF (5 mL)under N₂. The reaction mixture was heated at 110-120° C. for 16 h. Aftercooling, the reaction mixture was diluted with water (30 mL) andacidified with 1N HCl. The precipitate was collected by filtration,washed with water, and dried over P₂O₅ under vacuum to give yieldcompound 8 (1.06 g). This crude material was used in subsequentreactions without further purification.

Synthesis of 5-Methyl-2-mercapto benzothiazole

5-Methyl-2-mercapto benzothiazole (10): To a suspension of2-chloro-5-methyl aniline (9, 1.00 g, 7.06 mmol) and K₂CO₃ (2.80 g, 28.2mmol) in DMF (20 mL) was added CS₂ (0.50 mL, 8.50 mmol) under N₂ at roomtemperature. The reaction mixture was heated at 150° C. for 16 h. Aftercooling, the reaction mixture was diluted with water (30 mL) andacidified with 1N HCl. The precipitate was collected by filtration,washed with water, and dried over P₂O₅ under vacuum to yield compound 10(0.35 g, 27%).

Synthesis of 5-Nitro-2-mercapto benzothiazole

5-Nitro-2-mercapto benzothiazole (12): A mixture of 2-fluoro-5-nitroaniline (11, 1.00 g, 6.40 mmol) and potassium O-ethyl dithiocarbonate(1.53 g, 9.60 mmol) in DMF (15 mL) was heated overnight at 110-110° C.under N₂. After cooling, the reaction mixture was diluted with water andacidified with 1N HCl. The precipitate was collected by filtration,washed with water, and dried under vacuum to yield compound 12 (1.27 g,94%).

Synthesis of 5-Cyano-2-mercaptobenzothiazole

5-Cyano-2-mercaptobenzothiazole (14): A mixture of 3-amino-4-chlorobenzonitrile (13, 0.50 g, 3.28 mmol) and potassium O-ethyldithiocarbonate (1.05 g, 6.55 mmol) in DMF (15 mL) was heated overnightat 110-120° C. under N₂. After cooling, the reaction mixture was dilutedwith water and acidified with 1N HCl. The precipitate was collected byfiltration, washed with water, and dried under vacuum to yield 14 (0.48g, 76%).

General Procedure A: Nucleophilic displacement of □-chloro ketones

To a solution of a substituted 2-mercaptobenzothiazole (16.5 mmol) indry THF (50 mL) at 0° C. was added NaH (60% in mineral oil, 18.2 mmol)in small portions. After stirring for 30 min at 0° C., a solution ofcompound 2 or 4 (16.5 mmol) in THF (10 mL) was added dropwise. Thereaction was stirred overnight at room temperature. The reaction mixturewas quenched by addition of water (25 mL) and EtOAc (50 mL), the twolayers were separated. The aqueous layer was extracted with EtOAc (2×20mL). All the combined organics were washed with water (2×20 mL), brine(20 mL), and dried over MgSO₄. The solvent was removed under reducedpressure and the product was purified by column chromatography to affordthe desired product.

2-(5-Chlorobenzothiazol-2-ylsulfanyl)-1-(8-methyl-3,4-dihydro-2H-quinolin-1-yl)-ethanone(15): The title compound was synthesized according to General ProcedureA using compound 2 as the electrophile and5-chloro-2-mercaptobenzothiazole as the nucleophile to yield 15 (3.60 g,56%): MS m/z: 393 (M+H)⁺. Anal. Calcd. For, C₁₉H₁₇ClN₂OS₂.0.2H₂O: C,58.14; H, 4.47; N, 7.14; S, 16.33; Cl, 9.03. Found: C, 57.90; H, 4.40;N, 7.17; S, 16.47; Cl, 9.24.

2-(1H-Benzoimidazol-2-ylsulfanyl)-1-(3,4-dihydro-2H-quinolin-1-yl)-ethanone(16): The title compound was synthesized according to General ProcedureA using compound 4 as the electrophile and 1H-benzo[d]imidazole-2-thiolas the nucleophile to yield 16 as a white solid (82%): MS m/z: 324(M+H)⁺. Anal. Calcd. For, C₁₈H₁₇N₃OS: C, 66.85; H, 5.30; N, 12.99; S,9.91. Found: C, 66.78; H, 5.41; N, 12.90; S, 9.72.

1-(3,4-Dihydro-2H-quinolin-1-yl)-2-(1-methyl-1H-benzoimidazol-2-ylsulfanyl)ethanone(17): The title compound was synthesized according to General ProcedureA using compound 4 as the electrophile and1-methyl-1H-benzo[d]imidazole-2-thiol as the nucleophile to yield 17 asa pale yellow solid (77%): MS m/z: 338 (M+H)⁺. Anal. Calcd. For,C₁₉H₁₉N₃OS: C, 67.63; H, 5.68; N, 12.45; S, 9.50. Found: C, 67.45; H,5.74; N, 12.25; S, 9.41.

1-(3,4-Dihydro-2H-quinolin-1-yl)-2-(5-methoxy-benzothiazol-2-ylsulfanyl)-ethanone(18):

The title compound was synthesized according to General Procedure Ausing compound 4 as the electrophile and5-methoxybenzo[d]thiazole-2-thiol as the nucleophile to yield 18 as apale yellow solid (83%): MS m/z: 371 (M+H)⁺. Anal. Calcd. For,C₁₉H₁₈N₂O₂S₂: C, 61.60; H, 4.90; N, 7.56; S, 17.31. Found: C, 61.40; H,4.90; N, 7.48; S, 17.26.

1-(3,4-Dihydro-2H-quinolin-1-yl)-2-(4,5-diphenyl-1H-imidazol-2-ylsulfanyl)-ethanone(19): The title compound was synthesized according to General ProcedureA using compound 4 as the electrophile and4,5-diphenyl-1H-imidazole-2-thiol as the nucleophile to yield 19 as aoff-white solid (26%): MS m/z: 426 (M+H)⁺. Anal. Calcd. For,C₂₆H₂₃N₃OS.0.2H₂O: C, 72.77; H, 5.50; N, 9.79; S, 7.47. Found: C, 72.58;H, 5.33; N, 9.74; S, 7.48.

1-(3,4-Dihydro-2H-quinolin-1-yl)-2-(4,5-diphenyl-1H-oxazol-2-ylsulfanyl)-ethanone(20): The title compound was synthesized according to General ProcedureA using compound 4 as the electrophile and 4,5-diphenyloxazole-2-thiolas the nucleophile to yield 20 as an off-white solid (40%): MS m/z: 427(M+H)⁺. Anal. Calcd. For, C₂₆H₂₂N₂O₂S: C, 73.21; H, 5.20; N, 6.57; S,7.52. Found: C, 73.49; H, 5.13; N, 6.57; S, 7.42.

1-(3,4-Dihydro-2H-quinolin-1-yl)-2-(6-nitro-benzothiazol-2-ylsulfanyl)-ethanone(21): The title compound was synthesized according to General ProcedureA using compound 4 as the electrophile and6-nitrobenzo[d]thiazole-2-thiol as the nucleophile to yield 21 as ayellow solid (33%): MS m/z: 386 (M+H)⁺. Anal. Calcd. For, C₁₈H₁₅N₃O₃S₂:C, 56.09; H, 3.92; N, 10.90; S, 16.64. Found: C, 55.98; H, 3.75; N,10.94; S, 16.41.

2-(4-Chloro-benzothiazol-2-ylsulfanyl)-1-(3,4-dihydro-2H-quinolin-1-yl)-ethanone(22): The title compound was synthesized according to General ProcedureA using compound 4 as the electrophile and4-chlorobenzo[d]thiazole-2-thiol as the nucleophile to yield 22 as awhite solid (72%): MS m/z: 375 (M+H)⁺. Anal. Calcd. For, C₁₈H₁₅ClN₂OS₂:C, 57.67; H, 4.03; N, 7.47; S, 17.11; Cl, 9.46. Found: C, 57.79; H,4.08; N, 7.48; S, 17.33; Cl, 9.41.

1-(3,4-Dihydro-2H-quinolin-1-yl)-2-(5-trifluoromethyl-benzothiazol-2-ylsulfanyl)-ethanone(23): The title compound was synthesized according to General ProcedureA using compound 4 as the electrophile and5-(trifluoromethyl)benzo[d]thiazole-2-thiol as the nucleophile to yield23 as a white solid (80%): MS m/z: 409 (M+H)⁺. Anal. Calcd. For,C₁₉H₁₅F₃N₂OS₂: C, 55.87; H, 3.70; N, 6.86; S, 15.70; F, 13.95. Found: C,55.79; H, 3.71; N, 6.92; S, 15.67; F, 14.07.

1-(3,4-Dihydro-2H-quinolin-1-yl)-2-(1-phenyl-4-thiophen-2-yl-1H-imidzol-2-ylsulfanyl)-ethanone(24): The title compound was synthesized according to General ProcedureA using compound 4 as the electrophile and1-phenyl-4-(thiophen-2-yl)-1H-imidazole-2-thiol as the nucleophile toyield 24 as an off-white solid (47%): MS m/z: 432 (M+H)⁺. Anal. Calcd.For, C₂₄H₂₁N₃OS₂: C, 66.79; H, 4.90; N, 9.74; S, 14.86. Found: C, 66.74;H, 4.78; N, 9.65; S, 15.05.

2-(6-Chloro-benzothiazol-2-ylsulfanyl)-1-(3,4-dihydro-2H-quinolin-1-yl)-ethanone(25): The title compound was synthesized according to General ProcedureA using compound 4 as the electrophile and6-chlorobenzo[d]thiazole-2-thiol as the nucleophile to yield 25 as awhite solid (73%): MS m/z: 375 (M+H)⁺. Anal. Calcd. For, C₁₈H₁₅ClN₂OS₂:C, 57.67; H, 4.03; N, 7.47; S, 17.11; Cl, 9.46. Found: C, 57.50; H,4.05; N, 7.48; S, 16.86; Cl, 9.74.

2-(7-Chloro-benzothiazol-2-ylsulfanyl)-1-(3,4-dihydro-2H-quinolin-1-yl)-ethanone(26): The title compound was synthesized according to General ProcedureA using compound 4 as the electrophile and7-chlorobenzo[d]thiazole-2-thiol as the nucleophile to yield 26 as awhite solid (46%): MS m/z: 375 (M+H)⁺. Anal. Calcd. For, C₁₈H₁₅ClN₂OS₂:C, 57.67; H, 4.03; N, 7.47; S, 17.11; Cl, 9.46. Found: C, 57.50; H,3.99; N, 7.47; S, 17.36; Cl, 9.52.

1-(3,4-Dihydro-2H-quinolin-1-yl)-2-(1-phenyl-1H-imidazol-2-ylsulfanyl)-ethanone(27): The title compound was synthesized according to General ProcedureA using compound 4 as the electrophile and 1-phenyl-1H-imidazole-2-thiolas the nucleophile to yield 27 as a yellow gum (69%): MS m/z: 350(M+H)⁺. Anal. Calcd. For, C₂₀H₁₉N₃OS: C, 68.74; H, 5.48; N, 12.02; S,9.18. Found: C, 68.47; H, 5.71; N, 11.75; S, 8.96.

1-(3,4-Dihydro-2H-quinolin-1-yl)-2-(1-methyl-5-phenyl-1H-imidazol-2-ylsulfanyl)-ethanone(28): The title compound was synthesized according to General ProcedureA using compound 4 as the electrophile and1-methyl-5-phenyl-1H-imidazole-2-thiol as the nucleophile to yield 28 asan off-white solid (42%): MS m/z: 364 (M+H)⁺. Anal. Calcd. For,C₂₁H₂₁N₃OS: C, 69.39; H, 5.82; N, 11.56; S, 8.82. Found: C, 69.29; H,5.83; N, 11.40; S, 8.83.

2-(5,6-Dichloro-benzoimidazol-2-ylsulfanyl)-1-(3,4-dihydro-2H-quinolin-1-yl)-ethanone(29): The title compound was synthesized according to General ProcedureA using compound 4 as the electrophile and compound 8 as the nucleophileto yield 29 as a white solid (41%): MS m/z: 409 (M+H)⁺. Anal. Calcd.For, C₁₈H₁₄Cl₂N₂OS₂: C, 52.81; H, 3.45; N, 6.84; S, 15.67. Found: C,52.75; H, 3.62; N, 6.80; S, 15.56.

1-(3,4-Dihydro-2H-quinolin-yl)-2-(5-methyl-benzoimidazol-2-ylsulfanyl)-ethanone(30): The title compound was synthesized according to General ProcedureA using compound 4 as the electrophile and5-methylbenzo[d]thiazole-2-thiol (10) as the nucleophile to yield 30 asa white solid (60%): MS m/z: 355 (M+H)⁺. Anal. Calcd. For, C₁₉H₁₈N₂OS₂:C, 64.38; H, 5.12; N, 7.90; S, 18.09. Found: C, 64.54; H, 5.22; N, 7.84;S, 17.88.

1-(3,4-Dihydro-2H-quinolin-1-yl)-2-(5-nitro-benzothiazol-2-ylsulfanyl)-ethanone(31): The title compound was synthesized according to General ProcedureA using compound 4 as the electrophile and5-nitrobenzo[d]thiazole-2-thiol (12) as the nucleophile to yield 31 as apale yellow solid (56%): MS m/z: 386 (M+H)⁺. Anal. Calcd. For,C₁₈H₁₅N₃O₃S₂: C, 56.09; H, 3.92; N, 10.90; S, 16.64. Found: C, 56.22; H,3.95; N, 10.81; S, 16.67.

2-[2-(3,4-Dihydro-2H-quinolin-2-yl)-2-oxo-ethylsfanyl]-benzothiazol-5-carbonitrile(32): The title compound was synthesized according to General ProcedureA using compound 4 as the electrophile and2-mercaptobenzo[d]thiazole-5-carbonitrile (14) as the nucleophile toyield 32 as a white solid (74%): MS m/z: 366 (M+H)⁺. Anal. Calcd. For,C₁₉H₁₅N₃OS₂: C, 62.44; H, 4.14; N, 11.50; S, 17.55. Found: C, 62.47; H,4.36; N, 11.35; S, 17.65.

1-(3,4-dihydroquinolin-1(2H)-yl)-2-(4,5-dimethylthiazol-2-ylthio)ethanone(33): The title compound was synthesized according to General ProcedureA using compound 4 as the electrophile and 4,5-dimethylthiazole-2-thiolas the nucleophile to yield 33 as a cream solid (690 mg, 74%). MS(APCI): m/z 319 [M+H]⁺. Anal. Calcd. for C₁₆H₁₈N₂OS₂: C, 60.34; H, 5.70;N, 8.80. Found: C, 60.51; H, 5.71; N, 8.72.

2-(4-tert-butylthiazol-2ylthio)-1-(3,4-dihydroquinolin-1(2H)-yl)ethanone(34): The title compound was synthesized according to General ProcedureA using compound 4 as the electrophile and 4-tert-butylthiazole-2-thiolas the nucleophile to yield 34 (37%): MS: m/z+347.2 [M+H]⁺. Anal. Calcd(found) for C₁₈H₂₂N₂OS₂: C, 62.39; (62.60), H, 6.40; (6.43), N, 8.08;(8.06), S, 18.50; (18.39).

Synthesis of1-(3,4-dihydroquinolin-1(2H)-yl)-2-(thiazolo[5,4-b]pyridin-2-ylthio)ethanone(36)

1-(3,4-dihydroquinolin-1(2H)-yl)-2-(thiazolo[5,4-b]pyridin-2-ylthio)ethanone(36): The mixture of compound 4 (418 mg, 2.0 mmol), compound 35 (338 mg,2.0 mmol) and K₂CO₃ (276 mg, 2.0 mmol) in MeCN (20 mL) was refluxed overnight under N₂. After cooling to room temperature, the mixture waspartitioned between EtOAc (100 mL) and water (50 mL). The organic layerwas separated, washed with water and brine, dried over Na₂SO₄ andconcentrated. The crude material was purified by flash chromatography onsilica gel eluting with EtOAc/hexanes (20-50%). The solid obtained wastriturated with ether (5 mL) and hexanes (20 mL) over night. Afterfiltration, compound 36 was obtained as a pale yellow solid (414 mg,61%). MS (APCI): m/z 342 [M+H]⁺. Anal. Calcd. for C₁₇H₁₅N₃OS₂: C, 59.80;H, 4.43; N, 12.31. Found: C, 59.80; H, 4.37; N, 12.16.

General Procedure B: Amide formation with EDCI-HOBt

To a solution of an acid (1.5 mmol) in DCM (25 mL) was added adisubstituted amine (1.5 mmol). The mixture was cooled in an ice-waterbath, then DIPEA (3.75 mmol) was added, followed by HOBt (1.5 mmol) andEDCI (1.5 mmol). The reaction mixture was stirred at room temperatureover night (or monitored by TLC). The mixture was diluted with DCM (25mL), washed with H₂O, citric acid (10% in H₂O), NaHCO₃ (sat.), driedover Na₂SO₄ and concentrated in vacuum. The crude product was purifiedby flash chromatography on silica gel eluting with EtOAc/hexanes(10-40%).

1-(2-(Benzo[d]thiazol-2-ylthio)acetyl)-1,2,3,4-tetrahydro-benzo[b]azepin-5-one(37): The title compound was synthesized according to General ProcedureB using 2-(benzo[d]thiazol-2-ylthio)acetic acid and1,2,3,4-tetrahydrobenzo[b]azepin-5-one (5) to yield 37 (160 mg, 29%): MS(APCI): m/z 369 [M+H]⁺. Anal. Calcd. for C₁₉H₁₆N₂O₂S₂: C, 61.93; H,4.38; N, 7.60. Found: C, 61.72; H, 4.31; N, 7.40.

2-(Benzo[d]thiazol-2-ylthio)-1-(6-methyl-3,4-dihydroquinolin-1(2H)-yl)ethanone(38): The title compound was synthesized according to General ProcedureB using 2-(benzo[d]thiazol-2-ylthio)acetic acid and6-methyl-1,2,3,4-tetrahydroquinoline to yield 38 (450.5 mg, 85%): MS(APCI): m/z 355 [M+H]⁺. Anal. Calcd. for C₁₉H₁₈N₂OS₂: C, 64.39; H, 5.12;N, 7.90. Found: C, 64.55; H, 5.01; N, 7.88.

2-(Benzo[d]thiazol-2-ylthio)-1-(6-methoxy-3,4-dihydroquinolin-1(2H)-yl)ethanone(39): The title compound was synthesized according to General ProcedureB using 2-(benzo[d]thiazol-2-ylthio)acetic acid and6-methoxy-1,2,3,4-tetrahydroquinoline to yield 39 (245 mg, 1.5 mmol). MS(APCI: m/z 371 [M+H]⁺. Anal. Calcd. for C₁₉H₁₈N₂O₂S₂: C, 61.60; H, 4.90;N, 7.56. Found: C, 61.49; H, 4.87; N, 7.59.

2-(Benzothiazol-2-ylsulfanyl)-1-(2-methyl-3,4-dihydro-2H-quinolin-1-yl)-ethanone(40): The title compound was synthesized according to General ProcedureB using 2-(benzo[d]thiazol-2-ylthio)acetic acid and2-methyl-1,2,3,4-tetrahydroquinoline to yield 40 as a pale yellow solid(0.32 g, 40%); MS m/z: 355 (M+H)⁺. Anal. Calcd. For, C₁₉H₁₈N₂OS₂: C,64.38; H, 5.12; N, 7.90; S, 18.09. Found: C, 64.38; H, 5.22; N, 7.85; S,17.93.

2-(Benzothiazol-2-ylsulfanyl)-1-(3,4-dihydro-2H-quinoxalin-1-yl)-ethanone(41): The title compound was synthesized according to General ProcedureB using 2-(benzo[d]thiazol-2-ylthio)acetic acid and1,2,3,4-tetrahydroquinoxaline to yield 41 as a white solid (51%) MS m/z:342 (M+H)⁺. Anal. Calcd. For, C₁₇H₁₅N₃OS₂.0.2H₂O: C, 59.18; H, 4.50; N,12.18; S, 18.52. Found: C, 59.32; H, 4.65; N, 12.18; S, 18.25.

2-(Benzothiazol-2-ylsulfanyl)-1-(5-fluoro-3,4-dihydro-isoindol-2-yl)-ethanone(42): The title compound was synthesized according to General ProcedureB using 2-(benzo[d]thiazol-2-ylthio)acetic acid and 5-fluoroisoindolineto yield 42 as a white solid (47%): MS m/z: 345 (M+H)⁺. Anal Calcd. For,C₁₇H₁₃FN₂OS₂.0.3H₂O: C, 58.37; H, 3.92; N, 8.01; S, 18.33. Found: C,58.15; H, 3.86; N, 7.92; S, 18.05.

2-(Benzothiazol-2-ylsulfanyl)-1-(6-fluoro-2,3-dihydro-indol-1-yl)-ethanone(43): The title compound was synthesized according to General ProcedureB using 2-(benzo[d]thiazol-2-ylthio)acetic acid and 6-fluoroindoline toyield 43 as a white solid (70%): MS m/z: 345 (M+H)⁺. Anal. Calcd. For,C₁₇H₁₃N₂FOS₂: C, 59.28; H, 3.80; N, 8.13; S, 18.62; F, 5.52. Found: C,59.51; H, 3.85; N, 8.13; S, 18.73; F, 5.37.

2-(Benzothiazol-2-ylsulfanyl)-1-(2,3-dihydro-benzo[1,4]oxazin-4-yl)-ethanone(44): The title compound was synthesized according to General ProcedureB using 2-(benzo[d]thiazol-2-ylthio)acetic acid and3,4-dihydro-2H-benzo[b][1,4]oxazine to yield 44 as a white solid (78%):MS m/z: 343 (M+H)⁺. Anal. Calcd. For, C₁₇H₁₄N₂O₂S₂: C, 59.63; H, 4.12;N, 8.18; S, 18.73. Found: C, 59.73; H, 4.17; N, 8.18; S, 18.76.

2-(Benzothiazol-2-ylsulfanyl)-1-(2,3,4,5-tetrahydro-benzo[b]-azepin-1-yl)-ethanone(45): The title compound was synthesized according to General ProcedureB using 2-(benzo[d]thiazol-2-ylthio)acetic acid and2,3,4,5-tetrahydro-1H-benzo[b]azepine (6) to yield 45 as a white solid(23%): MS m/z: 355 (M+H)⁺. Anal. Calcd. For, C₁₉H₁₈N₂OS₂: C, 64.38; H,5.12; N, 7.90; S, 18.09. Found: C, 64.53; H, 5.06; N, 7.83; S, 17.93.

2-(Benzothiazol-2-ylsulfanyl)-1-(8-methyl-3,4-dihydro-2H-quinolin-1-yl)-ethanone(46): The title compound was synthesized according to General ProcedureB using 2-(benzo[d]thiazol-2-ylthio)acetic acid and8-methyl-1,2,3,4-tetrahydroquinoline to yield 46 as a white solid (39%):MS m/z: 355 (M+H)⁺. Anal. Calcd. For, C₁₉H₁₈N₂OS₂: C, 64.38; H, 5.12; N,7.90; S, 18.09. Found: C, 64.37; H, 5.02; N, 7.74; S, 18.10.

2-(Benzothiazol-2-ylsulfanyl)-1-(3,4-dihydro-2H-quinolin-1-yl)-propan-1-one(47): The title compound was synthesized according to General ProcedureB using 2-(benzo[d]thiazol-2-ylthio)propanoic acid and1,2,3,4-tetrahydroquinoline to yield 47 as a yellow oil (34%): MS m/z:355 (M+H)⁺. Anal. Calcd. For, C₁₉H₁₈N₂OS₂: C, 64.38; H, 5.12; N, 7.90;S, 18.09. Found: C, 64.38; H, 5.26; N, 7.97; S, 18.08.

Synthesis of1-(4-Acetyl-3,4-dihydro-2H-quinoxalin-1-yl)-2-(benzothiazol-2-ylsulfanyl)-ethanone(48)

1-(4-Acetyl-3,4-dihydro-2H-quinoxalin-1-yl)-2-(benzothiazol-2-ylsulfanyl)-ethanone(48): To a 0° C. solution of compound 41 (0.41 g, 1.20 mmol) in dryCH₂Cl₂ (20 mL) were added Ac₂O (1.12 mL, 12.0 mmol) followed by dropwiseaddition of Et₃N (1.12 mL, 8.00 mmol), and DMAP (10 mg) The reaction wasstirred for 18 h under N₂ at room temperature. The reaction mixture wasdiluted with CH₂Cl₂ (20 mL), washed with saturated NaHCO₃ (10 mL), water(2×10 mL), and saturated NaCl (2×10 mL). Dried over MgSO₄ and filtered.The solvent was removed under reduced pressure and the product waspurified by column chromatography (CH₂Cl₂/EtOAc 1:1), and trituratedfrom CH₂Cl₂/hexanes to afford 48 as a yellow solid (0.24 g, 52%); MSm/z: 384 (M+H)⁺. Anal. Calcd. For, C₁₉H₁₇N₃O₂S₂.0.1H₂O: C, 59.23; H,4.92; N, 10.91; S, 16.64. Found: C, 58.84; H, 4.55; N, 10.55; S, 16.32.

General Procedure C: Nitro Reduction

A mixture of the aromatic nitro compound (1.40 g, 3.64 mmol) andSnCl₂.2H₂O (4.10 g, 18.2 mmol) in EtOH (40 mL) was heated to reflux for3 h. The reaction mixture was cooled to room temperature and quenchedwith saturated Na₂CO₃. The solids were filtered through celite. Most ofthe filtrate was evaporated, and extracted with EtOAc (3×50 mL). All thecombined organic extracts were washed with water (2×20 mL), and brine(20 mL). The organic solution was dried over MgSO₄, filtered. Thesolvent was removed under reduced pressure, and the product was purifiedby column chromatography to yield the desired amine.

2-(6-Amino-benzothiazol-2-ylsulfanyl)-1-(3,4-dihydro-2H-quinolin-1-yl)-ethanone(49): The title compound was made according to General Procedure C using1-(3,4-Dihydro-2H-quinolin-1-yl)-2-(6-nitro-benzothiazol-2-ylsulfanyl)-ethanone(21) and the product was purified by column chromatography (2% Et₃N inEtOAc/hexanes 1:1) to afford compound 49 as a pale yellow solid (0.79 g,61%); MS m/z: 356 (M+H)⁺. Anal. Calcd. For, C₁₈H₁₇N₃OS₂: C, 60.82; H,4.82; N, 11.82; S, 18.04. Found: C, 61.06; H, 4.92; N, 11.53; S, 17.56.

2-(5-Amino-benzothiazol-2-ylsulfanyl)-1-(3,4-dihydro-2H-quinolin-1-yl)-ethanone(50): The title compound was synthesized according to General ProcedureC using1-(3,4-Dihydro-2H-quinolin-1-yl)-2-(5-nitro-benzothiazol-2-ylsulfanyl)-ethanone(31) to yield 50 as an off-white foam (56%): MS m/z: 356 (M+H)⁺. Anal.Calcd. For, C₁₈H₁₇N₃OS₂: C, 60.82; H, 4.82; N, 11.82; S, 18.04. Found:C, 60.51; H, 4.90; N, 11.44; S, 17.69.

Synthesis ofN-{2-[2-(3,4-Dihydro-2H-quinolin-1-yl)-2-oxo-ethylsulfanyl]-benzothiazol-6-yl}-acetamide(51)

N-{2-[2-(3,4-Dihydro-2H-quinolin-1-yl)-2-oxo-ethylsulfanyl]-benzothiazol-6-yl}-acetamide(51): To a 0° C. solution of compound 49 (0.16 g, 0.45 mmol) in dryCH₂Cl₂ (5 mL) was added Ac₂O (0.25 mL, 2.70 mmol) followed by dropwiseaddition of Et₃N (0.25 mL, 1.80 mmol). The reaction was stirred for 2 hunder N₂. The reaction mixture was diluted with CH₂Cl₂ (40 mL), washedwith saturated NaHCO₃ (10 mL), water (2×10 mL), and saturated NaCl (2×10mL). Dried over MgSO₄ and filtered. The solvent was removed underreduced pressure and the product was purified by column chromatography(EtOAc/hexanes 3:7) to afford 51 as a white solid (154 mg, 86%); MS m/z:398 (M+H)⁺. Anal. Calcd. For, C₂₀H₁₉N₃O₂S₂: C, 60.43; H, 4.82; N, 10.57;S, 16.13. Found: C, 60.54; H, 4.91; N, 10.41; S, 15.87.

Synthesis ofN-{2-[2-(3,4-Dihydro-2H-quinolin-1-yl)-2-oxo-ethylsulfanyl]-benzothiazol-5-yl}-acetamide(52)

N-{2-[2-(3,4-Dihydro-2H-quinolin-1-yl)-2-oxo-ethylsulfanyl]-benzothiazol-5-yl}-acetamide(52): Compound 52 was synthesized by the same procedure as above toyield 52 as an off-white solid (53%): MS m/z: 398 (M+H)⁺. Anal. Calcd.For, C₂₀H₁₉N₃O₂S₂.0.1H₂O: C, 60.13; H, 4.85; N, 10.52; S, 16.05. Found:C, 59.94; H, 4.99; N, 10.35; S, 15.80.

Synthesis ofN-{2-[2-(3,4-Dihydro-2H-quinolin-1-yl)-2-oxo-ethylsulfanyl]-benzothiazol-6-yl}-methanesulfonamide(53)

N-{2-[2-(3,4-Dihydro-2H-quinolin-1-yl)-2-oxo-ethylsulfanyl]-benzothiazol-6-yl}-methanesulfonamide(53): To a 0° C. solution of compound 49 (170 mg, 0.48 mmol) in dryCH₂Cl₂ (5 mL) were added methanesulfonyl chloride (78 μL) followed bydropwise addition of Et₃N (0.13 mL). The reaction was stirred for 2.5 hunder N₂ at room temperature. The reaction mixture was diluted withCH₂Cl₂ (40 mL), washed with 1N HCl (10 mL), water (2×10 mL), andsaturated NaCl (2×10 mL). Dried over MgSO₄ and filtered. The solvent wasremoved under reduced pressure, and the product was purified by columnchromatography (EtOAc/hexanes 4:1) to afford a mixture of mono andbis-sulfonamides (215 mg). To a solution of the mixture of mono andbis-sulfonamides (200 mg) in 3 mL of MeOH and THF (2:1) was added NaOH(38 mg in 0.1 mL of H₂O). The reaction was stirred for 45 min. at roomtemperature. Most of the solvent was evaporated under vacuum, theresidue was dissolved in water (15 mL), and extracted with EtOAc. Thecombined organics were dried over MgSO₄ and filtered. The solvent wasremoved under reduced pressure, and the product was purified by columnchromatography (EtOAc/hexanes 2:1) to afford 53 as a white solid (120mg); MS m/z: 434 (M+H)⁺. Anal. Calcd. For, C₁₉H₁₉N₃O₃S₃.0.1H₂O: C,52.39; H, 4.86; N, 9.65; S, 22.08. Found: 52.06; H, 4.57; N, 9.45; S,21.80.

Synthesis ofN-{2-[2-(3,4-Dihydro-2H-quinolin-1-yl)-2-oxo-ethylsulfanyl]-benzothiazol-5-yl}-methanesulfonamide(54)

N-{2-[2-(3,4-Dihydro-2H-quinolin-1-yl)-2-oxo-ethylsulfanyl]-benzothiazol-5-yl}-methanesulfonamide(54): To a 0° C. solution of compound 50 (160 g, 0.45 mmol) in dryCH₂Cl₂ (5 mL) was added methanesulfonyl chloride (70 μL, 0.90 mmol))followed by dropwise addition of pyridine (100 μL, 1.40 mmol). Thereaction was stirred overnight under N₂. The reaction mixture wasdiluted with CH₂Cl₂ (40 mL), washed with 1N HCl (10 mL), water (2×10mL), and saturated NaCl (2×10 mL). Dried over MgSO₄ and filtered. Thesolvent was removed under reduced pressure and the product was purifiedby column chromatography (EtOAc/hexanes 1:1) to afford 54 as a paleyellow solid (117 mg, 60%); MS m/z: 434 (M+H)⁺. Anal. Calcd. For,C₁₉H₁₉N₃O₃S₂: C, 52.63; H, 4.42; N, 9.69; S, 22.19. Found: C, 52.69; H,4.49; N, 9.54; S, 22.11.

Synthesis ofN-{2-[2-(3,4-Dihydro-2H-quinolin-1-yl)-2-oxo-ethylsulfanyl]-benzothiazol-6-yl}-benzamide(55)

N-{2-[2-(3,4-Dihydro-2H-quinolin-1-yl)-2-oxo-ethylsulfanyl]-benzothiazol-6-yl}-benzamide(55): To a 0° C. solution of compound 50 (0.22 g, 0.62 mmol) in dryCH₂Cl₂ (6 mL) was added benzoyl chloride (0.10 mL, 0.93 mmol) followedby dropwise addition of Et₃N (0.26 mL, 1.86 mmol). The reaction wasstirred overnight under N₂ at room temperature. The reaction mixture waspoured onto ice-water, and extracted with CH₂Cl₂ (3×20 mL). The combinedorganics extracts were washed with water (2×10 mL), and saturated NaCl(2×10 mL). Dried over MgSO₄ and filtered. The solvent was removed underreduced pressure, and the product was purified by column chromatography(EtOAc/hexanes 1:1) to afford 55 as a white solid (132 mg, 46%); MS m/z:460 (M+H)⁺. Anal. Calcd. For, C₂₅H₂₁N₃O₂S₂: C, 65.33; H, 4.61; N, 9.14;S, 13.95. Found: C, 65.08; H, 4.64; N, 8.99; S, 13.69.

General Procedure D: Amide Formation with EDCI

To a mixture of acid (2.5 mmol), aniline (2.5 mmol) and DMAP (2.5 mmol)in DCM (15 mL) was added EDCI (2.5 mmol). The reaction mixture wasstirred at room temperature over night (or monitored by TLC). Themixture was diluted with DCM (25 mL), washed with H₂O, HCl (1N), NaOH(1N), dried over Na₂SO₄ and concentrated in vacuo. The crude product waspurified by flash chromatography on silica gel eluting withEtOAc/hexanes (10˜30%).

2-(4-Chloro-2-methylphenoxy)-1-(3,4-dihydroquinolin-1(2H)-yl)ethanone(56): The title compound was synthesized according to General ProcedureD using 2-(4-chloro-2-methylphenoxy)acetic acid and compound 3 to yieldcompound 56 (365 mg, 46%): MS (APCI): m/z 316 [M+H]⁺. Anal. Calcd. forC₁₈H₁₈ClNO₂: C, 68.46; H, 5.75; N, 4.44. Found: C, 68.41; H, 5.92; N,4.51.

2-(4-Chloro-2-methylphenoxy)-1-(3,4-dihydroquinolin-1(2H)-yl)propan-1-one(57): The title compound was synthesized according to General ProcedureD using 2-(4-chloro-2-methylphenoxy)propanoic acid and compound 3 toyield 57 (350 mg, 46%): MS (APCI): m/z 330 [M+H]⁺. Anal. Calcd. forC₁₉H₂₀ClNO₂: C, 69.19; H, 6.11; N, 4.25. Found: C, 69.09; H, 6.10; N,4.31.

Synthesis of2-(Benzo[d]thiazol-2-yloxy)-1-(3,4-dihydroquinolin-1(2H)-yl)ethanone(62)

Ethyl 2-(benzo[d]thiazol-2-yloxy)acetate (60): To a solution of 59 (1.1g, 10.6 mmol) in THF (100 mL) cooled in an ice-water bath was added NaH(60%, 465 mg, 11.6 mmol) portionwise under N₂. The mixture was thenstirred at room temperature for half an hour, cooled in an ice-waterbath and a solution of 58 (1.80 g, 10.6 mmol) in THF (10 mL) was addeddropwise. It was refluxed for 1 hour and then stirred at roomtemperature over night. After diluting with EtOAc (150 mL), the solutionwas washed with water (3×) and brine, dried and concentrated. The crudematerial was purified by flash chromatography on silica gel eluting withEtOAc/hexanes (0˜10%) to give 60 as a white solid (1.32 g, 53%).2-(Benzo[d]thiazol-2-yloxy)acetic acid (61): To a solution of 60 (1.32g, 5.6 mmol) in MeOH was added NaOH (3M, 75 mL, 0.22 mol) dropwise. Themixture was stirred at room temperature over night. It was then cooledin an ice-water bath and acidified to pH<2 with HCl (2N). The mixturewas then extracted with CHCl₃ (3×). The combined organic layers weredried over Na₂SO₄ and concentrated in vacuum to yield 61 as a whitesolid (1.0 g, 86%).2-(Benzo[d]thiazol-2-yloxy)-1-(3,4-dihydroquinolin-1(2H)-yl)ethanone(62): Compound 62 was synthesized according to General Procedure B usingcompound 3 and compound 61 to yield 62 (183 mg, 28%): MS (APCI): m/z 325[M+H]⁺. Anal. Calcd. for C₁₈H₁₆N₂O₂S: C, 66.64; H, 4.97; N, 8.64. Found:C, 66.88; H, 4.94; N, 8.60.

Synthesis of2-(benzo[d]thiazol-2-ylsulfinyl)-1-(3,4-dihydroquinolin-1(2H)-yl)ethanone(65) and2-(benzo[d]thiazol-2-ylsulfonyl)-1-(3,4-dihydroquinolin-1(2H)-yl)ethanone(66)

2-(Benzo[d]thiazol-2-ylthio)-1-(3,4-dihydroquinolin-1(2H)-yl)ethanone(64): Compound 64 was synthesized according to General Procedure B usingacid 63 and compound 3 to yield 64 (418 mg, 82%).2-(Benzo[d]thiazol-2-ylsulfinyl)-1-(3,4-dihydroquinolin-1(2H)-yl)ethanone(65): To a solution of compound 64 (167 mg, 0.49 mmol) in DCM (4 mL)cooled in a salt-ice bath was added a solution of mCPBA (77%, 110 mg,0.49 mmol) in DCM (4 mL) drop wise. The reaction mixture was stirred for1 hr. After diluting with DCM (30 mL), the solution was washed withNa₂S₂O₃, NaHCO₃, dried over Na₂SO₄ and concentrated in vacuum. The crudematerial was purified by flash chromatography on silica gel eluting withEtOAc/hexanes (20˜50%) to give compound 65 as a white solid (164 mg,94%). MS (APCI): m/z 357 [M+H]⁺. Anal. Calcd for C₁₈H₁₆N₂O₂S₂: C, 60.65;H, 4.52; N, 7.86. Found: C, 60.44; H, 4.66; N, 7.73.2-(Benzo[d]thiazol-2-ylsulfonyl)-1-(3,4-dihydroquinolin-1(2H)-yl)ethanone(66): To a solution of 64 (167 mg, 0.49 mmol) in DCM (4 mL) in anice-water bath was added a solution of mCPBA (77%, 412 mg, 1.84 mmol) inDCM (10 mL) drop wise. NaHCO₃ (154 mg, 1.84 mmol) was then added portionwise. The reaction mixture was stirred at room temperature over night.After diluting with DCM (50 mL), the solution was washed with H₂O,Na₂S₂O₃, NaHCO₃, dried over Na₂SO₄ and concentrated in vacuum. The crudematerial was purified by flash chromatography on silica gel eluting withEtOAc/hexanes (20˜40%) to give 66 as a white solid (157 mg, 86%). MS(APCI): m/z 373 [M+H]⁺. Anal. Calcd for C₁₈H₁₆N₂O₃S₂: C, 58.04; H, 4.33;N, 7.52. Found: C, 58.04; H, 4.43; N, 7.50.

Synthesis of1-(3,4-dihydroquinolin-1(2H)-yl)-2-(thiazolo[4,5-]pyridine-2-ylthio)ethanonehydrochloride (70)

Thiazolo[4,5-]pyridine-2-thiol (68): Compound 67 (1.0 g, 8.3 mmol) wasdissolved in 15 mL of dry DMF under N₂. KSCSOEt (2.0 g, 12.5 mmol) wasadded to the solution as a solid. The reaction mixture was stirred at95-100° C. for 15 h. TLC (70% EtOAc/Hex) showed consumption of startingmaterial. The reaction was quenched with ˜35 mL of H₂O followed byaddition of 40 mL of 1M HCl. The mixture was stirred for 30 min, duringwhich time precipitate was obtained. The precipitate was filtered,washed with 250 mL of H₂O and dried under vacuum over P₄O₁₀ to yieldcompound 68 (0.980 g, 70%).1-(3,4-Dihydroquinolin-1(2H)-yl)-2-(thiazolo[4,5-c]pyridin-2-ylthio)ethanone(69): Compound 68 (0.5 g, 3.0 mmol) was mixed with 20 mL of dry THF and20 mL of dry DMF under N₂ and the suspension was stirred for 30 min atRT and then cooled to 0° C. NaH (0.15 g 60% in mineral oil, 3.7 mmol)was added to the ice-cold suspension which became a greenish solutionupon stirring for several minutes at 0° C. The reaction mixture wasstirred for another 30 min at 0° C. and then2-chloro-1-(3,4-dihydroquinolin-1(2H)-yl)ethanone (0.63 g, 3.0 mmol)dissolved in 5 ml of dry THF was added. After the reaction mixture wasstirred for 16 h at RT (monitored by TLC (CH₂Cl₂/EtOAc=1:1 and MS), itwas diluted with 100 mL of EtOAc followed by addition of 50 mL of H₂O.The organic layer was separated and the aqueous layer was extracted withEtOAc (2×50 mL). The organic layers were combined washed with water,brine, dried over Na₂SO₄ and concentrated giving 1.1 g of crude whichwas purified by flash chromatography (CH₂Cl₂/EtOAc) to yield compound 69(0.5 g, 49%). MS: m/z+342.1 [M+H]⁺; Anal. Calcd (found) for C₁₇H₁₅N₃OS₂:C, 59.80; (59.89), H, 4.43; (4.51), N, 12.31; (12.12), S, 18.78;(18.53).

1-(3,4-dihydroquinolin-1(2H)-yl)-2-(thiazolo[4,5-c]pyridine-2-ylthio)ethanonehydrochloride (70): Compound 69 (162 mg, 4.7 mmol) was dissolved in 10mL of Et₂O and 2.5 mL of EtOH, stirred for 10 min, then 0.57 mL of 1MHCl (in Et₂O) was added slowly. The precipitation started upon additionof the acid. The reaction mixture was stirred for 1 h at RT, 40 min at0° C., filtered. The precipitate was washed with Et₂O and dried underhigh vacuum over P₄O₁₀ overnight, then at 45° C. for 4 h to yieldcompound 70 (150 mg, 77%): MS: m/z+342.1 (M+H)⁺; Anal. Calcd (found) forC₁₇H₁₆ClN₃OS₂: C, 54.03; (53.78), H, 4.27; (4.33), N, 11.12; (10.95),Cl, 9.38; (9.15), S, 16.97; (16.68).

Synthesis of2-(Benzo[b]thiophen-2-ylthio)-1-(3,4-dihydroquinolin-1(2H)-yl)ethanone(73)

Benzo[b]thiophene-2-thiol (72): Compound 71 (1.75 g, 13.0 mmol) wasdissolved in 40 mL of dry Et₂O under N₂ and the solution was cooled to40° C. nBuLi (2.5 M solution in hexane, 5.7 mL, 14.3 mmol) was added andthe reaction mixture was allowed to warm up to 0° C. during 1 h and thensulfur (0.414 g, 13.0 mmol) was added in one portion. The solution wasallowed to slowly warm up to RT over 3 h. Excess of 10% HCl was addedand the reaction mixture was stirred for 20 min. The thiol was extractedwith ether, washed with brine, dried over Na₂SO₄ and concentrated toafford 2.0 g of crude compound 72 which was used in the next stepwithout further purification.2-(Benzo[b]thiophen-2-ylthio)-1-(3,4-dihydroquinolin-1(2H)-yl)ethanone(73): Crude compound 72 (0.71 g, 4.3 mmol) was dissolved in 40 mL of dryTHF under N₂, stirred for 30 min at RT and then cooled down to 0° C. NaH(60% in mineral oil, 0.224 g, 5.5 mmol) was added to the ice-coldsolution which became a yellowish suspension. The formed suspension wasstirred for 40 min at 0° C. and then2-chloro-1-(3,4-dihydroquinolin-1(2H)-yl)ethanone (0.89 g, 4.3 mmol) wasadded as a solution in 4 mL of dry THF. The reaction mixture was stirredat RT for 16 h. The work-up, the same as for compound 5, gave crudematerial which was purified by flash chromatography (CH₂Cl₂/EtOAc)giving 0.56 g of the crude product. Pumping the crude product under highvacuum over P₄O₁₀ at 45° C. for 4 h removed the co-crystallized EtOAc toyield the desired product 73 (36%): MS: m/z⁺ 340.1 [M+H]⁺; Anal. Calcd(found) for C₁₉H₁₇NOS₂: C, 67.22; (67.18), H, 5.05; (5.11), N, 4.13;(4.08), S, 18.89; (18.70).

Synthesis of1-(3,4-dihydroquinolin-1(2H)-yl)-2-(3-methylbenzo[b]thiophen-2-ylthio)-ethanone(76)

3-Methylbenzo[b]thiophene-2-thiol (75): Compound 75 was obtainedaccording to the procedure used to synthesize compound 72, except theaddition of sulfur was done at 40° C. in an attempt to avoid significantformation of disulfide. The obtained crude material (75) was used forthe next step without further purification.1-(3,4-dihydroquinolin-1(2H)-yl)-2-(3-methylbenzo[b]thiophen-2-ylthio)-ethanone(76): Crude compound 75 (0.55 g, 3.0 mmol) was coupled with compound 4(0.48 g, 2.3 mmol) according to procedure used to synthesize compound73. The product was purified by column chromatography to yield compound76 (95 mg, 5% for two steps): MS: m/z+354.1 [M+H]⁺; Anal. Calcd (found)for C20H19NOS2.0.1H₂O: C, 67.61; (67.59), H, 5.45; (5.48), N, 3.94;(3.94), S, 18.05; (17.75).

Synthesis of2-(2-(3,4-dihydroquinolin-1(2H)-yl)-2-oxoethylthio)-1H-benzo[d]imidazole-1-carbonitrile(79)

2-(2-(3,4-Dihydroquinolin-1(2H)-yl)-2-oxoethylthio)-1H-benzo[d]imidazole-1-carbonitrile(79): To a solution of compound 16 (0.52 g, 1.6 mmol, 1.0 eq) in 6 mL ofdry THF stirred at −78° C. under N₂ LiHMDS (1.7 mL, 1.05 eq) was added.The mixture was stirred for 15 min. at −78° C. and then TosCN (306 mg,1.05 eq. in 1 mL of THF) was added dropwise. The reaction mixture wasquickly allowed to warmed up to RT. The reaction mixture was quenchedwith 10 mL of NH₄Cl (sat.), extracted into ethyl acetate, washed withbrine and concentrated to give creamy solid. The crude was purified bycolumn chromatography (ethyl acetate/hexanes) to give 79 (330 mg, 59%).MP=153.3-154.2° C. Anal/Calcd. for C₁₉H₁₆H₄OS: C, 65.50; H, 4.63; N,16.08; S, 9.20. Found C, 65.59; H, 4.66; N, 16.06, S 9.16.

Synthesis ofBenzo[d]thiazol-2-yl(2-(3,4-dihydroquinolin-1(2H)-yl)-2-oxoethyl)cyanamide(84)

Tert-butyl 2-(3,4-dihydroquinolin-1(2H)-yl)-2-oxoethylcarbamate (81): Toa solution of Boc Gly-OH (80) (4.0 g, 22.83 mmol) in dry CH₂Cl₂ (50 mL)at 0° C. under N₂ was added HATU (8.70 g, 22.9 mmol). After stirring for15 min at r.t., 1,2,3,4-tetrahydroquinoline (2.9 mL, 23.0 mmol), andDIPEA (12 mL, 68.8 mmol) were added. The reaction was stirred for 36 hat room temperature. The reaction mixture was diluted with CH₂Cl₂ (100mL) and washed with saturated NaHCO₃, 5% HCl, water, and saturated NaCl.Dried over MgSO₄, and filtered. The solvent was removed under reducedpressure and the product was purified by column chromatography (10-50%EtOAc in hexanes) to afford 81 as a solid (5.76 g, 87%).2-Amino-1-(3,4-dihydroquinolin-1(2H)-yl)ethanone, TFA salt (82):Compound 81 (2.8 g, 9.66 mmol) was dissolved in 20 mL of TFA/CH₂Cl₂(1:3) at 0° C. under N₂ After stirring for 1 h, the solvents wereremoved under reduced pressure. 5 mL of CH₂Cl₂/hexanes were added andremoved again under reduced pressure to afford 82 (3.0 g). This crudematerial was used in the next step with out further purification.2-(Benzo[d]thiazol-2-ylamino)-1-(3,4-dihydroquinolin-1(2H)-yl)ethanone(83): To a 0° C. solution of 2-chlorobenzothiazole (1.20 mL, 9.66 mmol)in 1-butanol (25 mL) was added compound 82 (2.94 g, 9.66 mmol)) followedby dropwise addition of Et₃N (4.5 mL, 32 mmol). The reaction was heatedto reflux for 24 h under N₂. The solvent was removed under reducedpressure and the product was purified by column chromatography (20-50%EtOAc in hexanes) to afford 83 as a white foam (2.58 g, 82%); MS m/z:324 (M+H)⁺. Anal. Calcd. For, C₁₈H₁₇N₃OS: C, 66.85; H, 5.30; N, 12.99;S, 9.91. Found: C, 67.01; H, 5.42; N, 12.78; S, 9.85.Benzo[d]thiazol-2-yl(2-(3,4-dihydroquinolin-1(2H)-yl)-2-oxoethyl)cyanamide(84): To a suspension of compound 83 (340 g, 1.05 mmol) in dry CH₃CN (10mL) were added cyanogen bromide (5M in CH₃CN) (0.42 mL, 2.1 mmol) andK₂CO₃ (580 g, 4.2 mmol). The reaction was heated to reflux for 4 h underN₂. 0.84 mL (4.2 mmol) of cyanogen bromide was added and the reflux wascontinued for 12 h. The reaction mixture was diluted with EtOAc (50 mL).The solids were filtered off, filtrate was concentrated, and the yellowresidue was purified by column chromatography (20% EtOAc in hexanes) toafford 84 as an off-white solid (80 mg, 28%). MS m/z: 349 (M+H)⁺. Anal.Calcd. For, C₁₉H₁₆N₄OS.0.6H₂O: C, 63.53; H, 4.83; N, 15.60; S, 8.92.Found: C, 63.24; H, 4.67; N, 15.25; S, 8.76.

Example 2 High-Throughput Screening Assay

The assay depends on detection of the rise in intracellular Ca²⁺concentration ([Ca²⁺]_(i)) following channel activation in cellsinducibly expressing the TRPV3 channel. Ca²⁺ rise is quantified with theuse of fluorescent Ca²⁺ indicators that are loaded into cells andthereafter indicated the [Ca²⁺]_(i). Ca²⁺ influx follows activation ofthe TRPV3 channel. Compounds inhibiting this [Ca²⁺]_(i) rise areconsidered hits for further investigation.

The commercially available HEK293/x line (Invitrogen) was stablytransfected with a TRPV3 construct and screened by immunostaining tofind clones with TRPV3 expression following stimulation with 1 μg/mltetracycline. Clonal TRPV3-expressing cells were maintained in thegrowth medium recommended by the manufacturer supplemented with 100μg/ml hygromycin to promote retention of the TRPV3 construct. Aftergrowing to near confluency, cells are plated at a density of ˜25,000cells/well in 384 well plates in the presence of 1 μg/ml tetracycline,and allowed to grow for 20-30 hrs. A nearly confluent monolayer results.Cells are then loaded with Ca²⁺ dye: Fura-2/AM or Fluo4/AM are added tothe wells to a final concentration of 2 μM or 1 μM, respectively, andincubated for 80 min or 60 min, respectively, at room temperature.Supernatant is then removed from the cells by inverting plates with asharp flick, and 40 μl Ringer's solution (140 mM NaCl, 4.5 mM KCl, 2 mMCaCl₂, 1 mM MgCl₂, 10 mM HEPES, 10 mM glucose, pH 7.4) is then added toeach well. Following ˜1 hour for recovery from loading, cells areassayed using the Hamamatsu FDSS 6000 system, which permits illuminationalternately at 340 nM and 380 nM for Fura-2 experiments, or at 485 nMfor Fluo4 experiments. Frames were acquired at a rate of 0.2 Hz. Duringthe assay, the plates are continuously vortexed, with pipette mixing ofwells following addition of each reagent. For the screening assay, 13 μlof a diluted stock of each compound to be tested (at 50 μM) was added toeach well for 2 minutes following the collection of a short (4 frame)baseline. 13 μl 750 μM 2-APB (2-aminoethyldiphenylborinate) was added toeach well, achieving a final concentration of 10 μM each compound and150 μM 2-APB. Data were collected for ˜3 minutes following addition of2-APB, where the fluorescent intensity (for Fluo4) and the F340/F380ratio (for Fura-2) are proportional to the [Ca²⁺]_(i). Negative controlsconsisted of HEK293/TREx TRPV3 cells exposed to 2-APB, but no testcompound. Positive control cells were usually HEK293/TREx (“parental”)cells exposed to 2-APB but no test compound, but sometimes normalHEK/293 TREx TRPV3 cells were also used, but not exposed to 2-APB ortest compound. These controls defined a screening window, and “hits”were defined as those test compounds inhibiting the fluorescenceresponse by at least 40%.

Example 3 Patch Clamp Experiments

Whole-cell patch clamp experiments permit the detection of currentsthrough the TRPV3 channel in the cell line described above. A glasselectrode is brought into contact with a single cell and the membrane isthen ruptured, permitting control of the voltage of the cell membraneand measurement of currents flowing across the membrane using theamplifier attached to the electrode. A perfusion system permits controlof the extracellular solution, including the addition of blockers andactivators of the current. The current can be activated by heating thissolution to 28° C. or warmer or by addition of 20 μM 2-APB to thesolution.

TRPV3 cells were induced 2048 hours, removed from growth plates, andreplated at low density (to attain good single-cell physical separation)on glass coverslips for measurement. In some cases, cells were grown inlow density overnight on glass coverslips. Patch clamp recordings weremade in the whole-cell mode with a holding potential of 40 mV. Every 5seconds, a voltage ramp was applied from −120 to +100 mV, 400 ms induration. Currents elicited were quantified at −80 mV and +80 mV. Theinternal solution consisted of 140 mM cesium aspartate, 10 mM EGTA, 2.27mM MgCl₂, 1.91 mM CaCl₂ and 10 mM HEPES, pH to 7.2 with KOH; with 50 nMcalculated free Ca²⁺. External solution was Ringer's solution describedabove. Upon addition of 2-APB or upon heating of the extraceullarsolution as described above, TRPV3 current was induced only inTRPV3-expressing cells and not in parental HEK293 TREx cells. Thiscurrent showed a small inward component, reversal near +10 mV and astrong outward rectification, and is referred to as Phase I. Uponcontinued or repeated readdition of 2-APB or heat as a stimulus, currentcharacteristics change, resulting in a Phase II that is linear through+10 mV. Removal of the stimulus caused most of the current to go away,and inhibitor addition could still inhibit this current.

FIGS. 14-17 summarize data collected for various tested compounds. Thedata includes approximate IC₅₀ values for inhibition of TRPV3 mediatedinward and outward current (phase 1 and phase 2) as assessed bypatch-clamp, as well as comparative data for inhibition of other ionchannels. Note that several of the compounds inhibit at least onefunction of TRPV3 with an IC₅₀ of 1000 nM (1 uM) or less. Thesecompounds include at least one small molecule that inhibits a functionof TRPV3 with an IC₅₀ of 500 nM or less. Note that all IC₅₀ valuesprovided in FIGS. 14-17 are given in nM.

To determine whether compounds were selective for TRPV3 inhibition overinhibition of other ion channel types, the human ERG (hERG), NaV1.2, andTRPV1 (hTRPV1) channels and the rat TRPV6 (rTRPV6) channel were alsostably transfected and expressed or induced to express in mammalian celllines. The methods for measuring currents from these channels arewell-established and have been described in numerous publications (See,Weerapura et al., 2002, J Physiology 540: 15-27; Rush et al., 2005, JPhysiology 564: 808-815; Caterina et al., 1997, Nature 389: 816-824;Hoenderhop et al., 2001, J Physiology 537: 747-761; Clapham et al.,2003, Pharmacol Rev 55: 591-596). Compounds of interest were testedagainst these channels at concentrations up to 30 μM, and the resultingdata were used to estimate IC₅₀. Selectivity data for ruthenium red andfor selected compounds are provided in the following Table. Data ispresented as fold selective.

Fold Selectivity of Blockers for TRPV3 Over Other Channels CompoundTRPV3:hERG TRPV3:NaV1.2 TRPV3:TRPV1 TRPV3:TRPV6 Compound 64 >40 fold >40fold >40 fold >40 fold selective selective selective selective Compound15 >1000 fold >1000 fold >1000 fold >1000 fold selective selectiveselective selective Compound 46 >100 fold N/D >200 fold N/D selectiveselective Compound 101 >1000 fold N/D >1000 fold N/D selective selectiveCompound 102 >300 fold N/D N/D N/D selective Ruthenium >40 fold N/D 3fold selective 10 fold Red selective (Clapham et al.)

Example 4 Other Screening Assays

Although the exemplary TRPV3 inhibitors provided herein were identifiedusing the assays described in Examples 2 and 3, other cell-based assayscan be used to identify and/or characterize TRPV3 inhibitors. One suchassay is described in U.S. application Ser. No. 11,078,188, filed Mar.11, 2005, the contents of which are hereby incorporated by reference intheir entirety. TRPV3 protein can be expressed in the prokaryotic cellsystem described in application Ser. No. 11,078,188, and this system canbe used to screen for compounds that modulate an activity of the TRPV3protein. Alternatively, an ion channel other than TRPV3 can be expressedin the prokaryotic cell system, and the system can be used to evaluatethe activity profile of an identified TRPV3 inhibitors with respect toother ion channels.

Any assays performed to identify and/or characterize compounds thatinhibit an activity of TRPV3 can be performed in a high-throughputfashion, or can be performed on a smaller scale examining individualcompounds or small numbers of compounds. Additionally, any of theseassays can be performed (i) as a primary assay to identify compoundsthat inhibit a function of TRPV3; (ii) as a secondary assay to assessthe specificity of a compound with respect to its activity against otherion channels; (iii) as an assay used in a medicinal chemistry program tooptimize subject compounds.

Example 5 TRPV3 Antagonist Diminishes Pain from Thermal Injury

The thermal injury model was used to evaluate the effectiveness of anexemplary TRPV3 inhibitor in the treatment of nociceptive pain. FIG. 1provides a schematic representation of the protocol used to evaluate aTRPV3 inhibitor in the thermal injury model.

Briefly, the following protocol was followed. Male Holtzman rats(approximately 300 grams) were tested on thermal escape using aHargreaves type apparatus. Under light anesthesia, a thermal injury (52°C. for 45 seconds) was applied to one heel. The animals were tested forthermal escape latency of the injured and uninjured paw before and at30, 60, 80 and 120 minutes after injury. Drug (a TRPV3 inhibitor ormorphine) or vehicle (0.5% methylcellulose) was administered after thebaseline measurement and approximately 15-20 minutes prior to thethermal injury. In addition to the escape latency measurement,behavioral observations were made throughout the experiment.

The results of one series of experiments are summarized in FIG. 2. FIG.2 shows the effects of various concentrations of a TRPV3 inhibitor(inhibitor 64) on escape latency in the injured paw of treated micefollowing thermal injury, in comparison to a methylcellulose control.Note the significant sensitivity to pain in the vehicle injected injuredpaw, evidenced by the very fast (4 second) escape latency after injury.Increasing concentrations of the TRPV3 inhibitor significantlydiminished pain in the injured animals. This is evidenced by theincreasing escape latency measurements. In fact, administration of 200mg/kg of TRPV3 inhibitor resulted in an escape latency comparable tothat observed prior to injury. The reduction of pain observed in theinjured paw of animals injected with the TRPV3 inhibitor arestatistically significant.

The effects observed in the injured paw, and summarized in FIG. 2, arespecific to the injured paw. FIG. 3 summarizes experiments measuringescape latency in the uninjured paw. Even at the highest concentrationsadministered, TRPV3 inhibitor had no effect on the uninjured paw. Thisindicates that TRPV3 inhibitors may have fewer deleterious side-effectsthan other analgesic compounds. For example, and as shown in FIG. 4, thenarcotic morphine does have an effect on both the injured and un-injuredpaw.

Table A provides further data indicating that a TRPV3 inhibitor withefficacy in reducing pain following thermal injury, specificallyinhibitor 64, has few side effects then morphine.

TABLE A Pre Injury Behavioral deficits Observation t = 30 min t = 60 mint = 90 min t = 120 min TRPV3 Inhibitor 64 - Side Effects Observed atVarious Concentrations of Compound Spontaneous 0 of 8 (200 mg) 0 of 8(200 mg) 0 of 8 (200 mg) 0 of 8 (200 mg) 0 of 8(200 mg) agitation 0 of 8(50 mg) 0 of 8 (50 mg) 0 of 8 (50 mg) 0 of 8 (50 mg) 0 of 8 (50 mg) 0 of8 (12.5 mg) 0 of 8 (12.5 mg) 0 of 8 (12.5 mg) 0 of 8(12.5 mg) 0 of8(12.5 mg) Tactile allodynia 0 of 8 (200 mg) 0 of 8 (200 mg) 0 of 8 (200mg) 0 of 8 (200 mg) 0 of 8 (200 mg) 0 of 8 (50 mg) 0 of 8 (50 mg) 0 of 8(50 mg) 0 of 8 (50 mg) 0 of 8 (50 mg) 0 of 8 (12.5 mg) 0 of 8 (12.5 mg)0 of 8 (12.5 mg) 0 of 8(12.5 mg) 0 of 8 (12.5 mg) Pinna response 0 of 8(200 mg) 0 of 8 (200 mg) 0 of 8 (200 mg) 0 of 8 (200 mg) 0 of 8 (200 mg)0 of 8 (50 mg) 0 of 8 (50 mg) 0 of 8 (50 mg) 0 of 8 (50 mg) 0 of 8 (50mg) 0 of 8 (12.5 mg) 0 of 8 (12.5 mg) 0 of 8 (12.5 mg) 0 of 8(12.5 mg) 0of 8 (12.5 mg) Corneal response 0 of 8 (200 mg) 0 of 8 (200 mg) 0 of 8(200 mg) 0 of 8 (200 mg) 0 of 8 (200 mg) 0 of 8 (50 mg) 0 of 8 (50 mg) 0of 8 (50 mg) 0 of 8 (50 mg) 0 of 8 (50 mg) 0 of 8 (12.5 mg) 0 of 8 (12.5mg) 0 of 8 (12.5 mg) 0 of 8(12.5 mg) 0 of 8 (12.5 mg) Exopthalmos 0 of 8(200 mg) 0 of 8 (200 mg) 0 of 8 (200 mg) 0 of 8 (200 mg) 0 of 8 (200 mg)0 of 8 (50 mg) 0 of 8 (50 mg) 0 of 8 (50 mg) 0 of 8 (50 mg) 0 of 8 (50mg) 0 of 8 (12.5 mg) 0 of 8 (12.5 mg) 0 of 8 (12.5 mg) 0 of 8(12.5 mg) 0of 8 (12.5 mg) Catalepsy 0 of 8 (200 mg) 0 of 8 (200 mg) 0 of 8 (200 mg)0 of 8 (200 mg) 0 of 8 (200 mg) 0 of 8 (50 mg) 0 of 8 (50 mg) 0 of 8 (50mg) 0 of 8 (50 mg) 0 of 8 (50 mg) 0 of 8 (12.5 mg) 0 of 8 (12.5 mg) 0 of8 (12.5 mg) 0 of 8(12.5 mg) 0 of 8 (12.5 mg) Sedate 0 of 8 (200 mg) 0 of8 (200 mg) 2 of 8 (200 mg) 2 of 8 (200 mg) 2 of 8 (200 mg) 0 of 8 (50mg) 0 of 8 (50 mg) 0 of 8 (50 mg) 0 of 8 (50 mg) 0 of 8 (50 mg) 0 of 8(12.5 mg) 0 of 8 (12.5 mg) 0 of 8 (12.5 mg) 0 of 8(12.5 mg) 0 of 8 (12.5mg) Blistered paw 0 of 8 (200 mg) 0 of 8 (200 mg) 0 of 8 (200 mg) 1 of 8(200 mg) 1 of 8 (200 mg) 0 of 8 (50 mg) 0 of 8 (50 mg) 3 of 8 (50 mg) 4of 8 (50 mg) 5 of 8 (50 mg) 0 of 8 (12.5 mg) 0 of 8 (12.5 mg) 2 of 8(12.5 mg) 3 of 8(12.5 mg) 4 of 8 (12.5 mg) Morphine - Side EffectsObserved at Following Adminstration of 10 mg/kg Spontaneous 0 of 7 0 of7 0 of 7 0 of 7 0 of 7 agitation Tactile allodynia 0 of 7 0 of 7 0 of 70 of 7 0 of 7 Pinna response 0 of 7 5 of 7 2 of 7 0 of 7 0 of 7 Cornealresponse 0 of 7 0 of 7 0 of 7 0 of 7 0 of 7 Exopthalmos 0 of 7 6 of 7 6of 7 6 of 7 0 of 7 Catalepsy 0 of 7 4 of 7 3 of 7 1 of 7 0 of 7Blistered paw 0 of 7 0 of 7 0 of 7 0 of 7 0 of 7 0.5% Methycellulose -Administration of Control Spontaneous 0 of 8 0 of 8 0 of 8 0 of 8 0 of 8agitation Tactile allodynia 0 of 8 0 of 8 0 of 8 0 of 8 0 of 8 Pinnaresponse 0 of 8 0 of 8 0 of 8 0 of 8 0 of 8 Corneal response 0 of 8 0 of8 0 of 8 0 of 8 0 of 8 Catalepsy 0 of 8 0 of 8 0 of 8 0 of 8 0 of 8Blistered paw 0 of 8 0 of 8 6 of 8 7 of 8 7 of 8

Table A provides a comparison of the side-effects observed in ratsfollowing administration of TRPV3 inhibitor 64, at varyingconcentrations, versus the side-effects observed in rats administered 10mg/kg of morphine. Side-effects of rats administered 0.5%methycellulouse were observed and served as a control.

Briefly, some of the side-effects of morphine administration includeexopthalmos (bulging eyes), catalepsy (a near catatonic state), and thepinna response. These side-effects were not observed in ratsadministered 12.5, 50, or 200 mg/kg of TRPV3 inhibitor 64.

These experiments demonstrated that a TRPV3 inhibitor diminished pain inthe injured paw following thermal injury. The effectiveness of the TRPV3inhibitor was specific to the injured paw. This specificity indicatesthat TRPV3 inhibitors diminish pain without producing many of theside-effects of narcotics such as morphine. The efficacy of the TRPV3inhibitor in this model of nociceptive pain supports the use of TRPV3inhibitors in the treatment of severe pain, including chronic and acutepain.

Example 6 TRPV3 Antagonist Diminishes Pain following Formalin

As outlined above, the formalin model involves injection of a formalinsolution intradermally or intraperitoneally. Injection of formalinsolution invokes a biphasic response, and thus provides a model for bothnociceptive and inflammatory pain. The formalin model was used toevaluate the effectiveness of an exemplary TRPV3 inhibitor in thetreatment of pain. FIG. 5 provides a schematic representation of theprotocol used to evaluate a TRPV3 inhibitor in the formalin model.

Briefly, the following protocol was followed. Male Holtzmann ratsreceived intraplantar injections of 50 μL of 2% formalin. Paw flinchingwas detected by an automated sensor detecting movement of a small metalband placed on the injected hind paw. Drug or vehicle was administeredapproximately 15 minutes prior to the injection of formalin. Theanimal's response to injection of the irritant was measured by countingflinches per minutes during the Early Phase (the first 5 minutesfollowing injection of formalin), during the Late Phase (approximately30 minutes after injection of formalin), and during the intervening painfree phase.

The results of exemplary experiments are summarized in FIG. 6. Flinchesper minute, a measure of the pain and discomfort experienced by theanimal, was measured following formalin injection in animals receivingeither 200 mg/kg TRPV3 inhibitor or a vehicle control (0.5%methylcellulose). Administration of a TRPV3 inhibitor substantiallyreduced the flinches per minute during both Early Phase and Late Phase.These results indicated that a TRPV3 inhibitor diminished both phases ofpain in the formalin model. Note that these results are similar to thepublished data reporting the effectiveness of morphine in the formalinmodel. However, further experiments indicated that TRPV3 diminished painwithout producing any of the narcotic side-effects of morphine. Theefficacy of the TRPV3 inhibitor in both phases of this model ofnociceptive pain and inflammatory pain supports the use of TRPV3inhibitors in the treatment of severe pain, including chronic and acutepain.

Example 7 TRPV3 Antagonist Diminishes Pain in the Carrageenan Model ofAcute Inflammatory Pain

As outlined above, the carrageenan model is a model of acuteinflammatory pain. As such, it may be used to evaluate effectiveness inrelieving pain caused by inflammation, for example, pain due toarthritis. FIG. 7 provides a schematic representation of the protocolused to evaluate a TRPV3 inhibitor in the carrageenan model.

Briefly, naive rats were pretested for sensitivity to a heat stimulususing the Hargreaves apparatus. The next day, 100 μL of λ-carrageenanwas injected into the plantar surface of the right hindpaw approximately4.5 hours before testing. 30-60 minutes before testing the rats wereinjected intraperitoneally with vehicle or drug. In this study, the drugadministered was either a TRPV3 inhibitor or the TRPV1 inhibitor,A-425619 (produced by Abbott; maximal efficacious dose, 40 mg/kg IP; ElKouhen et al, 2005, J Pharmacol Exp Ther 314: 400-4091; Honore et al.,2005, J Pharmacol Exp Ther 314: 410-421). Following injection of bothcarrageenan and drug or vehicle control, the thermal escape latency wasmeasured. Data are expressed as the recorded Paw Withdrawal Latencies(PWLs) in seconds).

FIG. 8 summarizes the results of these experiments. For each “drug”(vehicle, TRPV3 inhibitor, TRPV1 inhibitor), the results of the controlpaw are shown at the left and the results of the experimental(carrageenan-injected) paw are shown at the right. The results show thatthe TRPV3 inhibitor diminished pain in the carrageenan model.Specifically, the TRPV3 inhibitor reduced thermal hyperalgesia in thismodel of inflammatory pain. At the higher tested concentration of TRPV3inhibitor (200 mg/kg), the TRPV3 inhibitor was as effective as the TRPV1inhibitor in diminishing pain. The efficacy of the TRPV3 inhibitor inthis model of inflammatory pain supports the use of TRPV3 inhibitors inthe treatment of inflammatory pain, for example, pain due to arthritis.

Example 8 TRPV3 Antagonists Diminish Pain in the CFA Model ofInflammatory Pain

As outlined above, the Complete Freund's Adjuvant (CFA) model is a modelof inflammatory pain. As such, it may be used to evaluate effectivenessin relieving pain caused by inflammation, for example, pain due toarthritis and other inflammatory conditions. FIG. 9 provides a schematicrepresentation of the protocol used to evaluate a TRPV3 inhibitor in theCFA model.

Naive rats were pretested for sensitivity to a heat stimulus in theHargreaves apparatus. The next day, 100 μL of complete Freund's adjuvant(CFA) was injected into the plantar surface of the right hindpaw. Twodays later, in the morning, the rats were again pretested. In theafternoon, rats were injected with either vehicle control(methylcellulose or PEG-400) or with drug (TRPV3 inhibitor 64, TRPV3inhibitor 15, TRPV1 inhibitor A-425619, or the non-steroidalanti-inflammatory diclofenac). Drugs or vehicle were injectedintraperitoneally, and 45 minutes later rats were tested forhyperalgesia by applying the heat source to the CFA injected anduninjected hindpaw and measuring latency to withdrawal.

FIG. 10 summarizes the results of experiments showing that the TRPV3inhibitor 64 reduced pain in the CFA model of inflammatory pain.Specifically, the TRPV3 inhibitor reduced thermal hyperalgesia in thismodel of inflammatory pain. The efficacy of this TRPV3 inhibitor wassimilar to that of diclofenac (a non-steroidal anti-inflammatory) and tothe TRPV1 inhibitor. In fact, at the highest dose of the TRPV3 inhibitor(200 mg/kg) tested, the effects of the TRPV3 inhibitor were more potentthan the effects of either the non-steroidal anti-inflammatory or theTRPV1 inhibitor.

FIG. 11 shows that the TRPV3 inhibitor 64 returns pain sensitivity tobaseline levels. Specifically, the TRPV3 inhibitor reduces pain in theinflamed paw back to where it was before the CFA was injected. Thecontrol paw (uninflamed) was not affected showing that the TRPV3antagonist does not prevent normal sensation (data not shown).

FIG. 12 summarizes the results of experiments showing that a secondTRPV3 inhibitor, 15, also reduces pain in the CFA model of inflammatorypain. Specifically, this second TRPV3 inhibitor reduced thermalhyperalgesia in the CFA model of inflammatory pain. The efficacy of thisTRPV3 inhibitor was similar to that of diclofenac (a non-steroidalanti-inflammatory). In fact, at the highest dose of the TRPV3 inhibitor(200 mg/kg) tested, the effects of the TRPV3 inhibitor were more potentthan the effects of the non-steroidal anti-inflammatory.

The efficacy of multiple TRPV3 inhibitors in this model of inflammatorypain supports the use of TRPV3 inhibitors in the treatment ofinflammatory pain, for example, pain due to arthritis. The tested TRPV3inhibitors appear to reduce pain without toxicity and without dullingnormal sensation (See, FIG. 11); the uninflamed paw was unaffected bytreatment with the TRPV3 antagonist (data not shown). Additionally, thetested TRPV3 inhibitors reduce pain with similar or greater efficacythan a non-steroidal anti-inflammatory or a TRPV1 inhibitor. Given theobserved side-effects of both of these classes of compounds, TRPV3inhibitors may reduce pain without the side-effects experienced withavailable analgesics.

Example 9 Plasma Levels of TRPV3 Inhibitors

The plasma levels of TRPV3 inhibitor 15 were determined via HPLC/MS/MSfollowing administration to male Sprague-Dawley rats. Rats wereadministered a single intravenous bolus dose of 1.2 mg/kg, anintraperitoneal dose of a suspension of 30 or 60 mg/kg, an oral dose ofa solution of 8.6 mg/kg, or an oral dose of a suspension of 6.9 mg/kg.

For intraveneous administration, TRPV3 inhibitor 15 solutions wereformulated in Solutol at a target concentration of 0.25 mg/mL. Thesolution was administered as a rapid bolus (approximately 2-3 seconds)via the tail vein of conscious rats or via jugular vein catheter inanesthetized rats. The solution was administered at a dose volume of 4mL/kg.

For intraperitoneal administration, TRPV3 inhibitor 15 was formulated asa uniform suspension in CMC (carboxymethylcellulose) at a targetconcentration of 5 mg/mL. The suspension was administered at a dosevolume of 10 mL/kg.

For oral administration, TRPV3 inhibitor 15 was formulated as either auniform suspension in CMC or as a solution in Solutol. The targetconcentration was 1 mg/mL. Oral administration was by gavage at a dosevolune of 10 mL/kg to conscious, fasted, male Sprague-Dawley rats.

FIG. 13 summarizes the results of experiments conducted followingintravenous administration (FIG. 13 a) or oral administration of asuspension (FIG. 13 b). Data for experiments conducted followingadministration via other route are not shown. Briefly, the estimatedhalf-life of TRPV3 inhibitor 15 is 1.6 hours. The estimated plasmaclearance was 41 mL/kg/min. The estimated volume of distribution was5510 mL/kg.

Estimated bioavailability in fasted rats was approximately 5%,regardless of whether a solution or suspension was administered. Theprofile of the plasma-concentration-time curve (FIG. 13) suggests thatTRPV3 inhibitor 15 was absorbed rapidly. This rapid absorption washighlighted by the observation that Cmax occurred after just 15 minutes.Such rapid absorption is an advantageous characteristic for a druguseful in the treatment of pain.

TRPV3 inhibitor 15 was prepared in formulations and administered viaseveral different routes of administration. This indicated that TRPV3inhibitors could be formulated in any of a number of ways and adapted tomost effectively treat particular diseases or injuries. Theseproperties, coupled to the minimal side-effects observed followingadministration of TRPV3 inhibitors to rats, indicated that TRPV3inhibitors have characteristics of suitable drugs and drug candidates.

Example 10 TRPV3 Inhibitors have a Unique Therapeutic Profile

Based on the foregoing experiments performed in multiple animal modelsof various types of pain, TRPV3 inhibitors have a unique profile incomparison to non-TRPV3 modulators used in the treatment of pain. Theunique profile of TRPV3 inhibitors, along with the absence of narcoticside-effects observed in these studies, makes TRPV3 inhibitorsparticularly well suited for use in the treatment of pain.

To briefly summarize, the foregoing experiments indicated that thetested TRPV3 antagonists are non-narcotic. The tested compounds wereeffective in an acute thermal injury model, a formalin model, thecarrageenan model, and the CFA model.

Table B provides a comparison, based on our data as well as dataavailable in the literature, between the test TRPV3 antagonists and thefollowing agents: morphine, COX-2 inhibitors (Vioxx® and Celebrex®),gabapentin, and a TRPV1 antagonist (A-425619)

In addition to the differences outlined in Table B, the non-TRPV3compounds have particular side-effects that must be considered beforeusing them for the treatment of pain. For example, morphine has narcoticand addictive effects that can impede daily functioning. COX-2inhibitors have been linked to an increased risk of cardiovascularcomplications. Gabapentin, also known as neurontin, was traditionallyused in the treatment of epilepsy. However, it has also been approvedfor and used in the treatment of post-herpetic neuralgia. Althougheffective, gabapentin does have numerous side-effects.

TABLE B TRPV3 COX-2 TRPV1 Antagonist Morphine Inhibitors GabapentinAntagonist Non-narcotic Non-narcotic Narcotic Non-narcotic Non-narcoticNon-narcotic Acute thermal Effective Effective Ineffective EffectiveEffective injury Formalin May be Effective in Effective in Effective inIneffective Model effective in both Phase I Phase II Phase II Phase Iresponse and response only response only response. Phase II Effective inresponse Phase II response Carrageenan Effective Effective EffectiveSlightly Effective Model effective CFA Model Effective EffectiveEffective Ineffective EffectiveIncorporation by Reference

All publications and patents mentioned herein, are hereby incorporatedby reference in their entirety as if each individual publication orpatent was specifically and individually indicated to be incorporated byreference.

Equivalents

Those skilled in the art will recognize, or be able to ascertain usingno more than routine experimentation, many equivalents to the specificembodiments of the invention described herein. Such equivalents areintended to be encompassed by the following claims.

1. A method for treating, reducing the frequency of, or delaying theonset of pain, comprising administering to a patient in need thereof aneffective amount of a compound of formula Va or a salt thereof:

wherein R₁ is absent or represents one or more substituents on the ringto which they are attached wherein each of said one or more substituentsare, independently for each occurrence, selected from among lower alkyl,alkoxy, carboxyl, amido, sulfonamido, heterocyclyl, cycloalkyl,hydroxyl, amino, acylamino, sulfonylamino, or R₁ taken together with thecarbon to which it is attached forms a carbonyl or thiocarbonyl, and R₂and R₃ are absent or represent one or more substituents on the ring towhich they are attached, wherein each of said one or more substituentsare, independently for each occurrence, selected from among lower alkyl,alkoxy, carboxyl, amido, sulfonamido, heterocyclyl, cycloalkyl,hydroxyl, amino, acylamino, sulfonylamino, nitro, halogen, CF₃, orcyano.
 2. The method of claim 1, wherein said compound inhibits at least95% of TRPV3-mediated current at 5 micromolar or less.
 3. The method ofclaim 1, wherein said compound inhibits TRPV3 with an IC₅₀ at least oneorder of magnitude lower than its IC₅₀ for inhibition of one or more ofTRPV5, TRPV6, NaV 1.2, mitochondrial uniporter and hERG channelactivities.
 4. The method of claim 1, wherein said compound inhibitsTRPV3 with an IC₅₀ of 100 nanomolar or less.
 5. The method of claim 1,wherein the compound is administered orally, transdermally,parenterally, rectally, vaginally, intranasally, intraocularly,intravenously, intramuscularly, intraarterially, intrathecally,intracapsularly, intraorbitally, intracardiacly, intradermally,intraperitoneally, transtracheally, subcutaneously, subcuticularly,intraarticularly, subcapsularly, subarachnoidly, intraspinally,intrasternally or by inhalation.
 6. The method of claim 1, wherein R₁ isabsent; R₂ represents H or CH₃ and is positioned ortho to the N of thering system; and R₃ represents one or more substituents positioned metaor para to the nitrogen of the ring system wherein said one or moresubstituents are selected from among lower alkyl, alkoxy, carboxyl,amido, sulfonamido, heterocyclyl, cyclo alkyl, hydroxyl, amino,acylamino, sulfonylamino, nitro, halogen, CF₃, or cyano.
 7. The methodof claim 1, wherein the compound is administered orally.
 8. The methodof claim 1, used to treat, reduce the frequency of, delay the onset of,or alleviate symptoms of acute pain, chronic pain, touch sensitivity,itching sensitivity, or as part of treating a burn.
 9. The method ofclaim 1, wherein R₁ is absent, or independently for each occurrence, isselected from lower alkyl, or alkoxy, or R₁ taken together with thecarbon to which it is attached forms a carbonyl; R₂ is absent, orindependently for each occurrence, is selected from lower alkyl, alkoxy,or halogen; and R₃ is absent, or independently for each occurrence, isselected from lower alkyl, alkoxy, CF₃, nitro, or halogen.
 10. Themethod of claim 8, wherein the pain is post-surgical pain, cancer pain,inflammatory pain, or neuropathic pain.